Polypeptides relating to signal transfer of advanced glycation end product receptor

ABSTRACT

It is intended to provide polypeptides which directly or indirectly bind to a cytoplasmic domain of RAGE and inhibits the signal transduction from binding of a ligand to RAGE (the receptor for advanced glycation endproduct) through activation of NFκB. Typical examples thereof are polypeptides respectively having amino acid sequences represented by SEQ ID NOS:1 to 32, SEQ ID NOS:67 to 79 and SEQ ID NOS:80 to 86 in Sequence Listing or the amino acid sequence including deletion, substitution or addition of one to several amino acids. Also, polypeptides involved in the signal transduction of RAGE are provided. Moreover, a pharmaceutical for gene therapy using polynucleotides encoding these polypeptides: a method for screening a useful compound using these polypeptides: and pharmaceutical composition and diagnostic reagent are also provided.

TECHNICAL FIELD

[0001] The present invention relates to novel polypeptides. Inparticular, the present invention relates to novel polypeptides involvedin a signal transduction of the receptor for an advanced glycation endproduct (AGE) (hereinafter, the receptor is also referred to as “RAGE”)and a polynucleotide encoding the same.

BACKGROUND ART

[0002] A non-enzymatic reaction of glucose or another reducing sugarwith an amino group in a protein or in a lipid generates a Schiff base,and then an irreversible reaction thereof results in advanced glycationendproducts (AGE). AGE is produced and accumulated by aging orhyperglycemia, and it cause denaturation of proteins, production ofactive oxygen, and activation of cells and inflammation. Theaccumulation of AGE is suggested in various kinds of diseases includingdiabetes, Alzheimer's Disease, dialysis amyloidosis, and also in naturalaging.

[0003] A binding to a specific receptor is regarded as one of themechanisms of AGE's action. Several receptors for AGE are known. Amongthem, RAGE (the receptor for AGE: AGE receptor) is considered to be amain receptor. RAGE is a single transmembrane receptor cloned in 1992,having three immunoglobulin domains on the extracellular domain.Examples of ligands thereof include AGE, amphoterin, and β-amyloid.

[0004] It is known in the art that the binding of AGE to RAGE causesactivation of NFκB by an oxidative stress. However, a signaltransduction from RAGE through NFκB has not been revealed up to now.

[0005] If a molecule involved in such a pathway is identified, it may bepossible to block a signal transduction of RAGE and thereby, inhibit theonset of a disease such as diabetes, a diabetic complication,Alzheimer's Disease, dialysis amyloidosis, a cancer, a periodontaldisease, or aging, by (i) exogenously introducing the molecule, (ii)inhibiting the expression of the molecule by a small molecule, (iii)inhibiting the function of the molecule by a small molecule, or thelike,

DISCLOSURE OF THE INVENTION

[0006] That is, an object of the present invention is to obtain amolecule involved in a signal transduction of RAGE and to use themolecule for treatment of diseases such as those listed above or forscreening of pharmaceuticals. In particular, (i) a gene therapy in whichthe obtained gene is introduced into human body, (ii) screening orevaluation of pharmaceuticals in cells in which the obtained gene isintroduced, or the like can be exemplified.

[0007] For finding out a protein to interact with a cytoplasmic domainof RAGE, the inventors of the present invention have screened humankidney cDNA libraries using a yeast two-hybrid system. Consequently, 31polynucleotides predicted to bind to a cytoplasmic domain of RAGE havebeen obtained. Furthermore, seven polynucleotides have been obtained asa result of screening human brain cDNA libraries, and thus the presentinvention has been completed.

[0008] That is, a gist of the present invention resides in apolypeptide, which is a substantially purified polypeptide, and ischaracterized by:

[0009] (a) directly or indirectly binds to a cytoplasmic domain ofreceptor for an advanced glycation end product (AGE); and

[0010] (b) inhibits a signal transduction from binding of a ligand toreceptor for the advanced glycation endproduct (AGE) through activationof NFκB.

[0011] According to a preferable embodiment of the above, there can bementioned an amino acid sequence selected from SEQ ID NOS: 1 to 32, SEQID NOS: 67 to 79, and SEQ ID NOS: 80 to 86 in the Sequence Listing orthe amino acid sequence including deletion, substitution, or addition ofone or several amino acids. According to a further preferable embodimentof the present invention, there can be mentioned an amino acid sequenceselected from SEQ ID NOS: 11, 12, 29, and 30 in the Sequence Listing orthe amino acid sequence including deletion, substitution, or addition ofone or several amino acids.

[0012] As a second gist of the present invention, a polynucleotide whichencodes the above-described polypeptide can be mentioned. According to apreferable embodiment thereof, there can be mentioned a nucleotidesequence represented by one of SEQ ID NOS: 33 to 63 and SEQ ID NOS: 87to 93 or a nucleotide sequence hybridizable with the polynucleotiderepresented by the nucleotide sequence under a stringent condition.According to a further preferable embodiment, there can be mentioned anucleotide sequence represented by one of SEQ ID NOS: 43, 44, 61, and 62or a nucleotide sequence hybridizable with the polynucleotiderepresented by the nucleotide sequence under a stringent condition.

[0013] Further, as a third gist of the present invention, there can bementioned a pharmaceutical for a gene therapy, comprising a vector whichis expressible in an animal and contains the above-describedpolynucleotide. According to a preferable embodiment thereof, there canbe mentioned a pharmaceutical for treating diabetes, a diabeticcomplication, Alzheimer's Disease, dialysis amyloidosis, cancer,periodontal disease, or an aging-related disease.

[0014] Further, as another gist of the present invention, there can bementioned a vector containing the above-mentioned polynucleotide; amicroorganism or cell which is transformed with the vector; and a methodfor producing the polypeptide, comprising culturing the microorganism orcell and isolating the polypeptide from the culture.

[0015] As a fourth gist of the present invention, there can be mentioneda method for screening a substance that inhibits or accelerates thebiding of the above-mentioned polypeptide to a cytoplasmic domain of thereceptor for AGE, comprising placing a target of screening in ascreening system that contains a cytoplasmic domain of the receptor forAGE and the polypeptide and measuring a degree of inhibition oracceleration of the binding of the polypeptide to the cytoplasmic domainof the receptor for AGE.

[0016] As a screening method, there can be mentioned as another gist ofthe present invention, a method of screening a substance that enhancesor inhibits a function of the polypeptide or a substance that increasesor decreases an amount of the polypeptide, comprising placing a targetof screening in a screening system that contains the above-mentionedpolypeptides and measuring a degree of enhancement or inhibition of thefunction of the polypeptide or a degree of increase or decrease in theamount of the polypeptide.

[0017] As a fifth gist of the present invention, there can be mentioneda compound which is obtainable by the screening method described above,and according to a preferable embodiment thereof, there can be mentioneda compound characterized by:

[0018] (a) directly or indirectly binds to a cytoplasmic domain of thereceptor for an advanced glycation endproduct (AGE); and

[0019] (b) inhibits a signal transduction from binding of a ligand withthe receptor for advanced glycation endproduct (AGE) through activationof NFκB.

[0020] Further, as another gist of the present invention, there can bementioned a pharmaceutical composition comprising, as an effectiveingredient, the substance selected from the group consisting of theabove-mentioned compound, a salt thereof, a hydrate thereof, and asolvate thereof. According to a preferable embodiment, thepharmaceutical composition may be used for treating a disease selectedfrom diabetes, a diabetic complication, Alzheimer's Disease, dialysisamyloidosis, a cancer, a periodontal disease, and an aging-relateddisease.

[0021] Further, as another gist of the present invention, there can bementioned an antibody which can specifically bind to the polypeptide; apolynucleotide which is represented by a nucleotide sequence having atleast 20 sequential nucleotides in any of the sequences of SEQ ID NOS:33 to 63 and SEQ ID NOS: 87 to 93, preferably in any of the sequences ofSEQ ID NOS: 43, 44, 61, and 62 in the Sequence Listing, a sequencecomplementary to the nucleotide sequence, and a nucleotide sequencehybridizable with the nucleotide sequence under a stringent condition; aprobe comprising the polynucleotide and a label; and a diagnosticreagent including the probe. According to a preferable embodiment, thediagnostic reagent may be used for a diagnosis of diabetes, a diabeticcomplication, Alzheimer's Disease, dialysis amyloidosis, a cancer, aperiodontal disease, and an aging-related disease.

BRIEF DESCRIPTION OF THE DRAWING

[0022]FIG. 1 is a diagram that illustrates functions of the clonedpolypeptides.

BEST MODE FOR CARRYING OUT THE INVENTION

[0023] Hereinafter, the present invention is described specifically. Thepolypeptide of the present invention is a substantially pure polypeptidewhich can directly or indirectly bind to a cytoplasmic domain of RAGEand inhibit signal transduction from binding of a ligand to RAGE throughactivation of NFκB.

[0024] The “direct biding” referred to herein means a direct binding ofthe polypeptide of the present invention to a cytoplasmic domain ofRAGE, and the “indirect biding” means a binding of the polypeptide ofthe present invention to a cytoplasmic domain of RAGE through a proteinor the like preexisting in an environment where the polypeptide of thepresent invention can bind to a cytoplasmic domain of RAGE (in an assaysystem or a human body, etc.). As “ligand”, there can be mentioned AGE,amphoterin, β-amyloid, or the like. For instance, when the polypeptideof the present invention is used for a method of screeningpharmaceuticals or for a gene therapy which will be described below, theligand can be selected as appropriate depending on a target disease. Forexample, AGE can be selected when the target disease is diabetes or adiabetic complication, amphoterin can be selected when the targetdisease is a cancer, and β-amyloid can be selected when the targetdisease is Alzheimer's Disease.

[0025] In the present invention, the “polypeptides” includes thosetypically recognized as peptides, oligopeptides, polypeptides, andproteins in the art. Natural proteins and chemically-synthesized orrecombinantly-engineered polypeptides and peptides are also included.The polypeptide may or may not be subjected to a post-translationalmodification such as glycosylation or phosphorylation.

[0026] A precursor of the polypeptide of the present invention is alsoincluded in the polypeptide of the present invention as far as it hasthe physiological activities as described above. Examples of such aprecursor include one having one or more amino acids added to theN-terminus and (or) the C-terminus of the peptide of the presentinvention.

[0027] Furthermore, the polypeptide of the present invention may bebound to polyethylene glycol for prolonging the half-period thereof, orfused with a secretory sequence or a leader sequence for secretion.Alternatively, the polypeptide of the present invention may be producedas a fusion polypeptide for purification.

[0028] A fragment characterized by the structural or functionalproperties of the polypeptide of the present invention is also useful.

[0029] A physiologically acceptable acid-added salt of the polypeptideof the present invention or of a precursor of the polypeptide is alsoincluded in the present invention. As a acid-added salt, there can bementioned a salt with an inorganic acid such as hydrochloric acid,phosphoric acid, or sulfuric acid; and a salt with an organic acid suchas acetic acid, formic acid, fumaric acid, maleic acid, succinic acid,citric acid, tartaric acid, malic acid, benzoic acid, or benzenesulfonicacid.

[0030] The “amino acid sequence including deletion, substitution ordeletion of one or several amino acids” of the present invention meansan amino acid sequence of a mutant of an allelic mutation or a mutationfound in nature, or an artificial mutation or a mutation obtainable byusing recombinant technology. All of the amino acid sequences to beincluded in the present invention are polypeptides having the sameactivity as that of the novel polypeptide molecule of the presentinvention. Even if only one amino acid residue is modified, an aminoacid sequence including variation which causes the loss of the activityis not included in the present invention.

[0031] The “polynucleotide” of the present invention is a polypeptidethat encodes the polypeptide of the present invention as describedabove. Specifically, the polynucleotide of the present invention is apolynucleotide encoding an amino acid sequence represented by one of SEQID NOS: 1 to 32, SEQ ID NOS: 67 to 79, and SEQ ID NOS: 80 to 86,preferably one of SEQ ID NOS: 11, 12, 29, and 30 in the SequenceListing. Specifically, such a polynucleotide can be exemplified by apolynucleotide having a nucleotide sequence represented by one of SEQ IDNOS: 33 to 63 and SEQ ID NOS: 87 to 93, preferably one of SEQ ID NOS:43, 44, 61, and 62. However, as far as the same amino acid sequence isencoded, each codon may be substituted with other equivalent codon.

[0032] The polynucleotide of the present invention may be apolynucleotide encoding a polypeptide having substantially the sameamino acid sequence as the amino acid sequence represented by one of SEQID NOS: 1 to 32, SEQ ID NOS: 67 to 79, and SEQ ID NOS: 80 to 86,preferably one of SEQ ID NOS: 11, 12, 29, and 30. Such a polynucleotideis preferably a polynucleotide having at least 80% homology to thenucleotide sequence represented by one of SEQ ID NOS: 33 to 63 and SEQID NOS: 87 to 93, preferably one of SEQ ID NOS: 43, 44, 61, and 62, orits complementary nucleotide sequence. A polynucleotide having 90%homology is more preferable, and in particular, the most preferable is apolynucleotide having 95% or more homology. As a polynucleotide having95% or more, preferably 97% or more homology, there can be mentioned apolynucleotide hybridizable with the polynucleotide represented by oneof SEQ ID NOS: 33 to 63 and SEQ ID NOS: 87 to 93, preferably one of SEQID NOS: 43, 44, 61, and 62 or a probe prepared from the sequence under astringent condition.

[0033] Furthermore, the present invention includes a polynucleotide thatencodes a fragment characterized by the structural or functionalproperties of the polypeptide of the present invention. Such apolynucleotide fragment and a polynucleotide hybridizable with apolynucleotide that encodes the fragment are useful as PCR primers aswell as probes for detecting a DNA that encodes the peptide of thepresent invention.

[0034] Hereinafter, the characteristic features included in the presentinvention are described with reference to Examples described below as arepresentative example. However, the present invention is not limited tothese examples.

[0035] For the purpose of obtaining a novel molecule involved in thesignal transduction from binding of AGE to RAGE through activation ofNFκB, a gene for a cytoplasmic domain of RAGE was amplified by a PCRmethod, as shown in Example 1 described later. Then, using the gene fora cytoplasmic domain of RAGE, human kidney cDNA libraries and brain cDNAlibraries were screened by a yeast two-hybrid method using the LexAsystem. Consequently, so far as the inventor of the present inventionknows, a polynucleotide was obtained, which encodes a novel polypeptidehaving no homology to any protein that has been reported so far.

[0036] In this way, 31 polynucleotides and 7 polynucleotides predictedto bind to a cytoplasmic domain of RAGE were obtained. Their nucleotidesequences are represented by one of SEQ ID NOS: 33 to 63 and 87 to 93,preferably one of SEQ ID NOS: 43, 44, 61, and 62 in the SequenceListing.

[0037] The nucleotide sequence of the polynucleotide cloned as describedabove can be linked with a vector which is expressible in a host celland contains transcriptional control activities such as a promoter, anoperator, or an enhancer; a termination sequence; and other regulatorysequences for controlling the expression of RAGE.

[0038] A recombinant vector containing the polynucleotide of the presentinvention is used for the production of the polypeptide of the presentinvention. The vector of the present inventionto be used is one suitablefor retention, amplification, and expression of a polynucleotide in ahost cell. The cloned polynucleotide that encodes the polypeptide of thepresent invention can be inserted into a vector directly or after beingdigested with a restriction enzyme or after being linked to a linker. ADNA has a translation initiation codon (ATG) on its 5′-terminal side anda translation termination codon (TAA, TGA, or TAG) on its 3′-terminalside. A DNA is located downstream from a promoter in the expressionvector.

[0039] Examples of such a vector include: plasmids derived fromEscherichia coli (such as pBR322, pBR325, and pUC12), plasmids derivedfrom Bacillus subtilis (such as pUB110 and pC194), plasmids derived fromthe genus Streptomyces, and plasmids derived from the genus Salmonella;plasmids derived from yeast episomes and host chromosome elements (suchas YCp plasmids and PYAC plasmids), bacteriophages such as λ-phage andvectors derived from viruses such as Vaccinia virus, Adenovirus,Retrovirus, and Baculovirus. Many of those vectors are commerciallyavailable.

[0040] A DNA sequence in the recombinant vector is operably-linked to anappropriate expression control sequence (promoter). Examples of such apromoter include: phage λPL promoter and T7 promoter; Escherichia colilac, trp, 1pp, and c promoters; Bacillus subtilis SPO1 promoter and penPpromoter; yeast PHO5 promoter, PGK promoter, GAP promoter, ADH1promoter, SUC2 promoter, GAL4 promoter, and MFα promoter; insect cellpolyhedron promoter and P10 promoter; and animal cell SV40 early or latepromoter, retrovirus LTR promoter, CMV promoter, HSV-TK promoter, andmetallothionein promoter.

[0041] In general, the expression vector contains an expression controlregion to be regulated by a repressor-binding region, an enhancer, orthe like. In addition, the expression vector contains a selectionmarker. Examples of appropriate markers include a dihydrofolatereductase (dhfr) gene and a neomycin-resistance gene for eukaryoticcells, and a tetracycline or ampicillin resistance gene for bacterialcells. The dhfr gene provides a transformed cell with methotrexateresistance, while the neomycin-resistance gene provides a transformedcell with G418 resistance. When a host cell is a dhfr gene-deficient CHOcell and the dhfr gene is used as a selection marker, a transformant canbe selected in a thymidine-free medium. In this case, a resistant straincan be selected by culturing cells under a gradually increasingconcentration of methotrexate (MTX). Thus, a DNA that encodes thepeptide of the present invention is amplified in the cell simultaneouslywith the amplification of the dhfr gene, thereby a CHO (dhfr-) cell withhigh level expression can be obtained.

[0042] The recombinant vector of the present invention is constructed ina way that a signal sequence is added to the N terminus of the peptide,if required. Such a signal sequence may be: a PhoA or OmpA signalsequence or the like in the case of an Escherichia coli host; an Mfα orSUC2 signal sequence or the like in the case of a yeast host; and anα-interferon signal sequence or the like in the case of an animal host.

[0043] The present invention also relates to a host cell that harborsthe recombinant vector as described above. Examples of the host cellsinclude mammalian cells, plant cells, insect cells, yeast cells,eukaryotic cells such as Aspergius fungi, and prokaryotic cells suchbacterial cells. The recombinant vector of the present invention can beintroduced into the host cell by means of calcium-phosphatetransfection, electroporation, transduction, infection, or the like.

[0044] Under the control of the promoter as described above, the peptideof the present invention can be expressed in hosts such as mammaliancells, yeasts, and bacteria as described above.

[0045] Examples of a prokaryotic host include Escherichia coli, Bacillussubtilis, Salmonella, Pseudomonas, Streptomyces, and Staphylococcus.Examples of the yeasts include Saccharomyces cerevisiae,Schizosaccharomyces pombe, and Pichia pastoris.

[0046] The transformed prokaryotic host is proliferated, and in a caseof using a vector containing an inducible promoter, a temperature or achemical inducer can initiate the induction. Then, the cells arecultured in a liquid medium containing a carbon source (such as glucose,dextran, or soluble starch), a nitrogen source (such as ammonium salt,nitrate salt, peptone, casein, meat extract, or bean cake), and aninorganic substance (such as calcium chloride, sodium dihydrogenphosphate, or magnesium chloride) at a suitable pH (i.e., at a pH ofabout 5 to 8) for a suitable period (i.e., for about 3 to 24 hours). Thesuitable culture temperature is about 14 to 43° C. for Escherichia coliand about 30 to 40° C. for bacteria of the genus Bacillus. The cell isphysically or chemically disrupted after cultivation, and then thepeptide of the present invention is purified from the resulting crudeextract.

[0047] The transformed yeast is cultured in a medium such as a minimalmedium at a pH of about 5 to 8 and at a temperature of about 20 to 35°C. for about 24 to 72 hours.

[0048] Insect cells to be used are Sf cells, MG1 cells, or the like whenthe virus is AcNPV (Autographa californica NPV), larval silkworms,silkworm cultured cells (BM-N cells) or the like when the virus isBombyx polynucleosis virus (BmPV). For the silkworm cells, a medium suchas a TC-10 medium containing a heat-inactivated 10% fetal bovine serumis used, and the cells are cultured to be confluent at about 27° C.,then the cells are subjected to passage.

[0049] Examples of the mammalian cell include COS-7 cell, mouse AtT-20cell, rat GH3 cell, rat MtT cell, mouse MIN6 cell, Vero cell, C127 cell,CHO cell, dhfr-gene defect CHO cell, HeLa cell, L cell, BHK cell,BALB3T3 cell, 293 cell, Bowes melanoma cell and the like.

[0050] Expression vectors for mammalian cell include a replicationorigin, a promoter (such as the above-mentioned SV40 early or latepromoter, retrovirus LTR promoter, CMV promoter, HSV-TK promoter, ormetallothionein promoter), an enhancer (such as SV40 enhancer,adenovirus enhancer, or cytomegalovirus early promoter), a selectionmarker (such as the above-mentioned dhfr gene or neomycin-resistancegene), a libosome binding site, a polyadenylation site (such as SV40polyadenylation site), splice donor and acceptor sites (such as a DNAsequence derived from SV40 splice site), a transcription terminationsequence, and a 5′-non-transcription sequence.

[0051] As such a vector, a plasmid vector, a single- or double-strandedphage vector, a single- or double-stranded RNA or DNA virus vector, orthe like can be used. As a medium for the transformed mammalian cells,an MEM medium, a DMEM medium, an RPMI-1640 medium, or the like, whichcontains a fetal bovine serum at a concentration of about 5 to 20%, canbe used. The culture of the cells is performed at a pH of about 6 to 8and a temperature of 30 to 40° C. for about 15 to 72 hours.

[0052] When the mammalian cells are used as hosts, the polypeptide ofthe present invention can be collected and purified from a culture ofthe recombinant cells using ammonium sulfate or ethanol precipitation,acid extraction, anion- or cation-exchange chromatography, hydrophobicinteraction chromatography, or the like.

[0053] The polypeptide of the present invention may not be glycosylated.In addition, depending on hosts, a polypeptide having methionine at itsN-terminus can be obtained.

[0054] Furthermore, the polynucleotide of the present invention isuseful in a gene therapy for treating a disease such as diabetes, adiabetic complication, Alzheimer's Disease, dialysis amyloidosis, acancer, a periodontal disease, or an aging-related disease. The “genetherapy” means the administration of a gene or gene-incorporated cellsinto the human body for treating a disease. For a pharmaceutical use,any of a purified polynucleotide, a recombinant, a culture liquor of atransformant, an isolated transformant, a treated-transformant, afixed-transformant, a crude enzyme solution, an enzyme-treated product,etc. may be used.

[0055] When the polynucleotide of the present invention is used for agene therapy, various technologies conventionally used for a genetherapy can be employed. Specifically, a gene containing thepolynucleotide of the present invention can be expressed in the body byimplanting the expression vector for the polynucleotide of the presentinvention, which is obtained from a virus vector such as a retrovirusvector, an adenovirus vector, an AAV vector, or a herpes virus vector orwhich is obtained by a membrane-fusion liposome method, into bone marrowcells of a patient suffering from a disease such as diabetes, a diabeticcomplication, Alzheimer's Disease, dialysis amyloidosis, a cancer, aperiodontal disease, or an aging-related disease (hereinafter, which maybe collectively referred to as “the disease”), according to a methodsuch as one described in JP 09-501046 A or a method based thereon; byadministering the expression vector into the muscle tissue, bloodsystem, intestine, lung, or the like of a patient suffering from thedisease according to a method such as one described in JP 09-505084 A ora method based thereon; or by administering the expression vector intothe cerebrospinal fluid of a patient suffering from the diseaseaccording to a method such as one described in JP 09-505561 A or amethod based thereon. Furthermore, an offspring of the patient can beprevented from suffering from the disease by incorporating the genecontaining the polynucleotide of the present invention into the egg cellof the patient suffering from the disease.

[0056] The polypeptide or polynucleotide of the present invention isuseful for identifying a compound which can modulate control of thesignal transduction from binding of a ligand to RAGE through activationof NFκB. That is, the present invention provides a method of screening asubstance that inhibits or accelerates a binding of the polypeptide ofthe present invention to a cytoplasmic domain of RAGE. The methodincludes: placing a target of screening in a screening system containinga cytoplasmic domain of RAGE and the polypeptide of the presentinvention; and measuring the degree of inhibition or acceleration of thebinding of the polypeptide of the present invention to a cytoplasmicdomain of RAGE.

[0057] In the present invention, the “target of screening” is notparticularly limited as far as it is a substance available for ascreening, the target may be a high molecule as well as a low molecule.For instance, the target of screening may be a peptide, an analogthereof, a microbial culture liquor, or an organic compound. In thepresent invention, “screening” is used in the meaning of including anassay.

[0058] The screening system is not limited as far as it is a methodtypically used in the art, for instance, an experimental system such asa yeast two-hybrid system can be used. That is, the interaction betweenthe polypeptide of the present invention and a cytoplasmic domain ofRAGE is monitored by the yeast two-hybrid activity, and then a substancethat inhibits or enhances the reporter activity thereof may be screed.In an alternative method, using the BIAcore (manufactured by BIACOREK.K.), a change in their interaction is monitored in the presence of asubstance to be provided as a screening target to identify the influenceof the substance on their interaction. In the screening describedherein, not only a polypeptide having the full-length polypeptide of thepresent invention having an amino acid sequence selected from SEQ IDNOS: 1 to 32, SEQ ID NOS: 67 to 79, and SEQ ID NOS: 80 to 86 in theSequence Listing but also a partial peptide having at least five aminoacid residues can be used as far as the target substance can be screenedin the above method. Preferably, it can be also possible to use not onlya polypeptide having the full-length polypeptide of the presentinvention having an amino acid sequence selected from SEQ ID NOS: 11,12, 29, and 30 but also a partial polypeptide having at least fiveresidues as far as the target substance can be screened in the abovemethod.

[0059] In the present invention, “measure the degree of inhibition ofbinding” is used in the meaning of including measuring the presence orabsence of the binding.

[0060] The methods as described above are methods in which a cytoplasmicdomain of RAGE is provided in the screening system. However, in thepresent invention, it is also possible to perform a method in which acytoplasmic domain of RAGE is not provided in a screening system. Thatis, there is provided a method of screening a substance that enhances orinhibits the function of the polypeptide of the present invention or asubstance that increases or decreases an amount of the polypeptide ofthe present invention, which method comprising: placing a targetsubstance in a screening system containing the polypeptide of thepresent invention; and measuring the degree of enhancement or inhibitionof the function of the polypeptide of the present invention or measuringthe degree of an increase or decrease in an amount of the polypeptide ofthe present invention.

[0061] The screening target, the definition of the screening, and thescreening system are as described above. “Measuring the degree ofenhancement or inhibition of the function” and “measuring the degree ofan increase or decrease in an amount” are used in the meaning ofmeasuring the presence or absence of enhancement or inhibition of thefunction and measuring the presence or absence of an increase ordecrease in the amount, respectively.

[0062] The screening system is not limited as far as it is a methodconventionally used in the art, for instance, a vector containing thepolypeptide of the present invention is introduced into a cell in whichpNFk-luc (STRATAGENE Co., Ltd.) is introduced; and then, a substancethat enhances or inhibits the NFκB activity is screened.

[0063] As described above, the polypeptide of the present invention isinvolved in the signal transduction from RAGE through NFκB, so that thescreening method of the present invention can be used as a method ofscreening a substance useful as a preventive agent and/or a therapeuticagent for treating a disease such as diabetes, a diabetic complication,Alzheimer's Disease, dialysis amyloidosis, a cancer, a periodontaldisease, or an aging-related disease.

[0064] The compounds obtainable by using the screening method of thepresent invention and the salts thereof, and their hydrates and solvatesare compounds that modify the binding of the polypeptide of the presentinvention to a cytoplasmic domain of RAGE (i.e., the compounds inhibitor accelerate the binding) or compounds having the cell-stimulatingactivities caused by the interaction between the polypeptide of thepresent invention and a cytoplasmic domain of RAGE. Examples of thecompounds include peptides, proteins, fermented products, non-peptidecompounds, and synthetic compounds. These compounds may be novel orknown compounds.

[0065] As a pharmaceutical composition, for example, the compound itselfmay be administered alone. It is preferable to prepare a pharmaceuticalcomposition containing the above compound as an effective ingredient byusing a pharmaceutically acceptable pharmaceutical additive, andadministrate the composition. The pharmaceutical composition may beorally or parenterally administered in a form of a pharmaceuticalcomposition or preparation (for example, a tablet, a pill agent, acapsule, a granule, powder, syrup, an emulsion agent, an elixir agent, asuspending agent, a resolvent, an injection, a drop, or a suppository)which is obtainable by mixing the compound of the present invention witha pharmaceutically acceptable carrier (for example, an excipient, abinder, a disintegrating agent, a flavoring substance, a deodorizingsubstance, an emulsifier, an attenuant, or a solubilizer). Thepharmaceutical composition can be manufactured according to theconventional method. In this specification, “parenterally” covers ahypodermic injection, an intravenous injection, an intramuscularinjection, an intraperitoneal injection, and a drip method. Acomposition for injection can be prepared by the method known in theart. A rectal suppository can be produced by mixing the pharmaceuticalcompound with a suitable excipient. Examples of solid formulation fororal administration include those as described above such as powder, agranule, a tablet, a pill agent, and a capsule. Examples of liquidformulations for oral administration include an emulsion agent, syrup,an elixir agent, a suspending agent, and a solution agent, each of whichis acceptable as a medicine.

[0066] Furthermore, each of the preparations as described above can beprepared by conventional procedures. The clinical dosage of thepharmaceutical of the present invention is determined by appropriatelyincreasing or decreasing on the basis of the substance used as aneffective ingredient, age, condition, symptom, the presence or absenceof simultaneous administration, and soon. The above-described dosage perday may be administered once a day or two or several times per day withappropriate intervals. Alternatively, the dosage may be intermittentlyadministered every several days.

[0067] On the basis of the nucleotide sequence information about thepolynucleotide of the present invention, it is possible to obtain anantisence specific to a polynucleotide complementary to the sequence.The “antisence” referred to herein means a polynucleotide complementaryto at least a part of mRNA or DNA that encodes the polynucleotide of thepresent invention, which inhibits the transcription and translation ofthe polynucleotide of the present invention. Furthermore, using thetransformant of the present invention, the effects of the antisence canbe also confirmed. For such an antisense, there can be used not onlytypical DNAs and RNAs but also any other sequence-specificnucleotide-like molecule considered to have antisence effects.

[0068] The polynucleotide and polypeptide of the present invention canbe used for diagnosing the above-described diseases. An antibodyspecific to the polypeptide of the present invention can be obtained byusing proteins which is produced in large amounts in a cell transformedby the polynucleotide of the present invention. As an immunizingantigen, the full-length polypeptide of the present invention or apeptide fragment consisting of at least five sequential amino acidresidues of the amino acid sequence of the polypeptide of the presentinvention may be used. In the present invention, the antibodies include,in addition to the complete antibodies, all of the molecules which areproduced from original antibody and considered to have antibody effects,for example, Fab, F(ab′)2, Fv, and scFv fragments that contain therespective antigen-binding sites.

[0069] Using the antibodies as described above, it is possible toconstruct the system of ELISA or RIA or the system of western blottingfor detecting the polypeptide of the present invention in cells andtissues. For instance, such a detecting system can be used fordiagnosing the diseases as described in the explanation of the screeningmethod.

[0070] Furthermore, in the present invention, the expression of thepolypeptide of the present invention in cells or tissues can be detectedusing a probe that contains a suitable label and the polynucleotiderepresented by a nucleotide sequence comprising at least 20 sequentialnucleotides in the sequence of any of SEQ ID NOS: 33 to 63 and SEQ IDNOS: 87 to 93, preferably any of SEQ ID NOS: 43, 44, 61, and 62 in theSequence Listing, a complementary sequence thereof, and a nucleotidesequence hybridizable with the nucleotide sequence under a stringentcondition. Therefore, such a probe can be also used in the assay fordiagnosis. A label used in this case is not particularly limited as faras it can be used in the conventional assay for the diagnosis. As such alabel, for example, avidin or biotin, an enzyme such as peroxidase, aradioactive isotope, a fluorescent substance, or an antigen can be used.Using those labels, the nucleotide sequences are labeled by a methodconventionally used in the art, and the labels can be detected bysuitable methods, respectively.

EXAMPLES

[0071] Hereinafter, the present invention will be described in moredetail with reference to the examples. However, the present invention isnot limited only to these examples.

Example 1

[0072] Cloning of a Protein Which Interacts With a Cytoplasmic Domain ofRAGE by Yeast Two-Hybrid Method

[0073] A gene for a cytoplasmic domain of RAGE (SEQ ID NO: 64 in theSequence Listing) (J. Biol. Chem Jul. 25, 1992 267 (21): 14998-5004) wasamplified using synthetic oligonucleotides:5′-ACGTGAATTCAGGCGGCAACGCCGAGGAG-3′ (SEQ ID NO: 65 in the SequenceListing) and 5′-CGATCTCGAGTCAAGGCCCTCCAGTACTACTC-3′ (SEQ ID NO: 66 inthe Sequence Listing), and was then digested with restriction enzymesEcoRI and XhoI. Also, a vector pHybLex/Zeo (manufactured by InvitrogenCo., Ltd.) was digested with the restriction enzymes EcoRI and XhoI.Subsequently, the both genes were subjected to a 14-hour ligationreaction using Takara Ligation Kit ver. 2 (manufactured by Takara ShuzoCo., Ltd.) and used for transforming E.coli strain DH5α (manufactured byTakara Shuzo Co., Ltd.), thereby colonies were obtained. Consequently, avector (pHybLex/Zeo-RAGECD) in which the cytoplasmic domain of RAGE isincorporated into the pHybLex/Zeo was obtained. Using a DNA sequencer(manufactured by Beckman Co., Ltd.), the sequence of the cytoplasmicdomain of RAGE was confirmed.

[0074] The vector pHybLex/Zeo-RAGECD obtained as described above wasintroduced into Yeast L40 strain attached to the MATCH MAKER kit whichis described below, and then, a clone into which the pHybLex/Zeo-RAGECDhad been introduced was selected.

[0075] Then, the Human kidney MATCHMAKER LexA library (manufactured byClontech Co., Ltd.) was introduced to the above-described strain and 10⁶colonies were screened. At the first screening, a histidine-deficientmedium was used and about 200 colonies were selected based on theexpression of a histidine gene, which was a reporter of the interaction.At the second screening, 31 colonies were obtained based on theexpression of LacZ, which was another reporter of the interaction.Finally, 31 clones each of which had been confirmed to surely have twokinds of reporter activities were selected. The above procedures werecarried out on the basis of protocols of the yeast two-hybrid methodfrom Invitrogen Co., Ltd. and Clontech Co., Ltd.

[0076] The library vectors were recovered from the yeast coloniesselected as described above, and the nucleotide sequences were analyzedusing the DNA sequencer (the same as above). The nucleotide sequencesare shown in SEQ ID NOS: 33 to 63 in the Sequence Listing.

[0077] The polypeptide obtained as described above (in a state of beinginserted in the pB42AD vector (manufactured by Clontech Co., Ltd.) bythe screening) was excised from the vector with the restriction enzymesEcoRI and XhoI. Then, the resulting fragments were subjected to aligation reaction, using the Ligation Kit ver. 2 (manufactured by TakaraShuzo Co., Ltd.), with the pGBKT7 vector (manufactured by Clontech Co.,Ltd.) which had been digested with the restriction enzymes EcoRI andSalI, thereby, the polypeptide-incorporated pGBKT7 vector was obtained.Similarly, the RAGECD fragments were excised from the above-describedpHybLex/Zeo-RAGECD vector with the restriction enzymes EcoRI and XhoI,followed by being ligated to a pGADGH vector (manufactured by ClontechCo., Ltd.) which had been digested with the restriction enzymes EcoRIand SalI, thereby a pGBKT7-RAGECD vector was obtained.

[0078] The pGBKT7-polypeptide vector was introduced into a yeast strainAH109 (manufactured by Clontech Co., Ltd.), and the pGADT7-RAGECD vectorwas introduced into a yeast strain Y187 (manufactured by Clontech Co.,Ltd.), by using the lithium acetate method (see page 20 of “YeastProtocols Handbook” attached to the kit of Clontech Co., Ltd.).

[0079] The plasmid-introduced yeasts were subjected to selection on atryptophan-deficient SD medium or on a leucine-deficient SD medium, andselected strains were then conjugated with the following combinations(see page 44 of “Yeast Protocols Handbook” attached to the kit ofClontech Co., Ltd.). Subsequently, the yeast in which two kinds ofplasmids had been introduced was selected.

[0080] 1. pGBKT7-polynucleotide of SEQ ID NO: 43+pGADT7-idle vector

[0081] 2. pGBKT7-polynucleotide of SEQ ID NO: 44+pGADT7-idle vector

[0082] 3. pGBKT7-polynucleotide of SEQ ID NO: 61+pGADT7-idle vector

[0083] 4. pGBKT7-polynucleotide of SEQ ID NO: 62+pGADT7-idle vector

[0084] 5. pGBKT7-polynucleotide of SEQ ID NO: 43+pGADT7-RAGECD vector

[0085] 6. pGBKT7-polynucleotide of SEQ ID NO: 44+pGADT7-RAGECD vector

[0086] 7. pGBKT7-polynucleotide of SEQ ID NO: 61+pGADT7-RAGECD vector

[0087] 8. pGBKT7-polynucleotide of SEQ ID NO: 62+pGADT7-RAGECD vector

[0088] As a result of the growth test of the above strains on the SDmedium lacking in tryptophan, leucine, histidine and adenine, strains 1,2, 3, and 4 were not grown, while strains 5, 6, 7, and 8 were grown.

[0089] Therefore, it was shown that the polynucleotides of SEQ ID NOS:43, 44, 61, and 62 were specifically bind to the RAGECD fragments,respectively.

[0090] That is, it was found that the polypeptide translated from eachof those polynucleotides binds to the cytoplasmic domain of RAGE.

[0091] The above procedures were carried out on the basis of protocolsof MATCH MAKER GAL4 Yeast Two Hybrid System 3 (manufactured by ClontechCo., Ltd.).

Example 2

[0092] Cloning of Protein Which Can Interact With a Cytoplasmic Domainof RAGE by Yeast Two-Hybrid Method

[0093] A gene for a cytoplasmic domain for RAGE (SEQ ID NO: 65 in theSequence Listing) (J. Biol. Chem 1992 Jul. 25, 267 (21): 14998-5004) wasamplified using synthetic oligonucleotides:5′-ACGTGAATTCAGGCGGCAACGCCGAGGAG-3′ (SEQ ID NO: 66 in the SequenceListing) and 5′-CGATCTCGAGTCAAGGCCCTCCAGTACTACTC-3′ (SEQ ID NO: 67 inthe Sequence Listing), and was then digested with restriction enzymesEcoRI and XhoI. Also, a vector pGBKT7 (manufactured by Clontech Co.,Ltd.) was digested with restriction enzymes EcoRI and Sal I.Subsequently, the both genes were subjected to a 14-hour ligationreaction using the Takara Ligation Kit ver. 2 (manufactured by TakaraShuzo Co., Ltd.) and used to transform E. Coli strain DH5α (manufacturedby Takara Shuzo Co., Ltd.), thereby colonies were obtained.Consequently, a vector (pGBKT7-RAGECD) in which the cytoplasmic domainof RAGE is introduced in the pGBKT7 was obtained. Using a DNA sequencer(manufactured by Beckman Co., Ltd.), the sequence of the cytoplasmicdomain of RAGE was confirmed.

[0094] The vector pGBKT7-RAGECD obtained as described above wasintroduced into yeast strain AH109 attached to the kit MATCH MAKERdescribed below. Then, a clone into which the PGBKT7-RAGECD had beenintroduced was selected.

[0095] Then, Pretransformated MATCHMAKER library Human Brain(manufactured by Clontech Co., Ltd.) was introduced into the abovestrain and 10⁶ colonies were screened. At the first screening, ahistidine-deficient medium was used, and about 140 colonies wereselected based on the expression of a histidine gene, which was areporter of the interaction. At the second screening, 40 colonies wereselected on an adenine-deficient medium based on the expression ofadenine, which was another reporter of the interaction.

[0096] Finally, 7 clones each of which had been confirmed to surely havetwo kinds of reporter activities were selected, the library vectors wererecovered, and the nucleotide sequences were then analyzed using the DNAsequencer (the same as above). The nucleotide sequences are shown in SEQID NOS: 87 to 93 in the Sequence Listing.

[0097] The above procedures were carried out on the basis of protocolsof the yeast two-hybrid method from Invitrogen Co., Ltd. and ClontechCo., Ltd.

Example 3

[0098] Functional Analysis on the Cloned Polypeptide

[0099] Because the polypeptide of SEQ ID NO: 62 was a part of, proteinkinase C zeta (PKC zeta), the full-length PKC zeta was cloned in pcDNA3.1 (+) (manufactured by Invitrogen Co., Ltd.) using the PCR method, andpcDNA 3.1 (+)—PKC zeta vector was obtained.

[0100] Using TransIT-LT1 (manufactured by Takara Shuzo Co., Ltd.),pNFkB-luc (manufactured by Stratagene Co., Ltd) was introduced into ratC6 glioma cells (ATCC CCL-107). The cells were cultured in a medium withthe addition of geneticin (manufactured by Nacalai Tesque, Inc.) toobtain the drug-resistant cells. Thus, stably expressing cells wereobtained.

[0101] Using the TransIT-LT1 (manufactured by Takara Shuzo Co., Ltd.),pcDNA 3.1 (+) and pcDNA 3.1 (+)—PKC zeta plasmids were introduced intothe cells, respectively. After 48 hours, the medium was replaced with aserum-free DMEM medium (manufactured by SIGMA Co., Ltd.). After anadditional 12 hours, the cells were stimulated for 6 hours with carboxymethyl lysine (CML) which had been prepared by incubating 1.7 g of BSA(manufactured by SIGMA Co., Ltd.) and 0.36 g of glyoxylic acid(manufactured by OHSIGMA Co., Ltd.) at 36° C. for 24 hours in thepresence of a sodium cyano borohydride (manufactured by SIGMA Co., Ltd.)as a catalyst. After the stimulation, a luciferase assay was performedusing Bright-Glo (manufactured by Promega Co., Ltd.), in which the cellswere measured with an ARVOsx plate reader (manufactured by Wallac Co.,Ltd.). Consequently, it was found that the introduction of pcDNA 3.1(+)—PKC zeta significantly increased the NFkB activity (FIG. 1) ascompared to the introduction of an empty vector pcDNA 3.1.

[0102] In the case of diseases such as a diabetic complication in whichRAGE may be involved, the activation of NFkB by AGE was known in theart. However, the intermediate pathway thereof had been unclear.

[0103] From this experiment, it was found that the polypeptide capableof interacting with the intracellular domain of the RAGE increases NFkBactivation due to AGE stimulation. Therefore, it was suggested that thepolynucleotide of SEQ ID NO: 62 would mediate the downstream signal ofRAGE.

[0104] Industrial Applicability

[0105] According to the present invention, a novel polypeptide involvedin the signal transduction of RAGE is obtained. In addition, thepolynucleotide encoding the polypeptide of the present invention can beused as a genetic source to be applied in a gene therapy. Furthermore,the polypeptide of the present invention is capable of providing amethod of screening a substance useful for treating diseases involved inthe signal transduction from RAGE through activation of NFκB, apharmaceutical composition obtainable by the screening method, and adiagnostic agent for those diseases.

[0106] Note that, the present application has been filed claiming apriority of Japanese Patent Application No. 2001-219122.

1 93 1 8 PRT Homo sapiens 1 Val Val Asp Met Arg Arg Tyr Phe 1 5 2 132PRT Homo sapiens 2 Met Val Thr Ala Gly His Ala Cys Thr Lys Lys Tyr ThrPro Glu Gln 1 5 10 15 Val Ala Met Ala Thr Val Thr Ala Leu His Arg ThrVal Pro Ala Ala 20 25 30 Val Pro Gly Ile Cys Phe Leu Ser Gly Gly Met SerGlu Glu Asp Ala 35 40 45 Thr Leu Asn Leu Asn Ala Ile Asn Leu Cys Pro LeuPro Lys Pro Trp 50 55 60 Lys Leu Ser Phe Ser Tyr Gly Arg Ala Leu Gln AlaSer Ala Leu Ala 65 70 75 80 Ala Trp Gly Gly Lys Ala Ala Asn Lys Glu AlaThr Gln Glu Ala Phe 85 90 95 Met Lys Arg Ala Met Ala Asn Cys Gln Ala AlaLys Gly Gln Tyr Val 100 105 110 His Thr Gly Ser Ser Gly Ala Ala Ser ThrGln Ser Leu Phe Thr Ala 115 120 125 Cys Tyr Thr Tyr 130 3 5 PRT Homosapiens 3 Ile Ile Ile Gly Ser 1 5 4 5 PRT Homo sapiens 4 Ser Leu Phe LeuMet 1 5 5 128 PRT Homo sapiens 5 Leu Val Ala Leu Pro Ala Thr Pro Leu ThrPro Ala Phe Pro Pro Pro 1 5 10 15 Ser Phe Asp Gln Arg Ser Ala Glu ThrLeu Glu Val Arg Lys Leu Asp 20 25 30 Thr His Pro Asp Arg Thr Gly Val LeuArg Gly Val Ala Gly Gly Phe 35 40 45 Gly Leu Asp Glu Phe Pro Pro Lys CysArg Arg Ser Gln Ala Arg Ala 50 55 60 Gln Lys Gln Arg Met Gly Lys Ser MetGln His Ser Val Tyr Leu Ile 65 70 75 80 Leu Thr Arg Gly Asn His Lys HisArg Lys Pro Gln Leu Asp Met Asp 85 90 95 Tyr Arg Val Leu Glu Lys Ser ValPhe His Gly Val Cys Leu His Leu 100 105 110 Leu Thr Ser Gln Val Leu LysGln Lys Thr Lys Ala Met Ile Leu Met 115 120 125 6 25 PRT Homo sapiens 6Arg Lys Ile Asn Pro Leu Ile Lys Leu Ile Asn His Ser Phe Ile Asp 1 5 1015 Leu Pro Thr Pro Ser Asn Ile Ser Ala 20 25 7 19 PRT Homo sapiens 7 SerVal Arg Gly Ser Val Val Gly Ser Thr Leu Thr Pro Leu Ala Cys 1 5 10 15Pro Asp Pro 8 15 PRT Homo sapiens 8 Gln Leu Cys Trp Leu Asp Trp Leu ProCys Ser Trp Pro Arg Thr 1 5 10 15 9 5 PRT Homo sapiens 9 Ser Phe Gly SerLeu 1 5 10 7 PRT Homo sapiens 10 Arg Lys Asn Lys Leu Arg Gln 1 5 11 21PRT Homo sapiens 11 Leu Arg Glu Lys Met Thr Gly Tyr Ile Gln Ser Asp MetAla Lys Ser 1 5 10 15 Leu Lys Val Leu Met 20 12 132 PRT Homo sapiensmisc_feature (124)..(124) “Xaa” may be any amino acid 12 Arg Val Thr IleArg Lys Ser Lys Asn Ile Leu Phe Val Ile Thr Lys 1 5 10 15 Pro Asp ValTyr Lys Ser Pro Ala Ser Asp Thr Tyr Ile Val Phe Gly 20 25 30 Glu Ala LysIle Glu Asp Leu Ser Gln Gln Ala Gln Leu Ala Ala Ala 35 40 45 Glu Lys PheLys Val Gln Gly Glu Ala Val Ser Asn Ile Gln Glu Asn 50 55 60 Thr Gln ThrPro Thr Val Gln Glu Glu Ser Glu Glu Glu Glu Val Asp 65 70 75 80 Glu ThrGly Val Glu Val Lys Asp Ile Glu Leu Val Met Ser Gln Ala 85 90 95 Asn ValSer Arg Ala Lys Ala Val Arg Ala Leu Lys Asn Asn Ser Asn 100 105 110 AspIle Val Asn Ala Ile Met Glu Leu Thr Met Xaa Pro Tyr Gly Ser 115 120 125Asn Phe Phe Trp 130 13 23 PRT Homo sapiens 13 Lys Gly Lys Asn Asp TrpLeu Tyr Ser Val Arg Tyr Gly Lys Lys Ser 1 5 10 15 Gln Gly Val Asn ValAsn Asp 20 14 66 PRT Homo sapiens 14 Glu His Met Val Ile Thr Asp Arg IleGlu Asn Ile Asp His Leu Gly 1 5 10 15 Phe Phe Ile Tyr Arg Leu Cys HisAsp Lys Glu Thr Tyr Lys Leu Gln 20 25 30 Arg Arg Glu Thr Ile Lys Gly IleGln Lys Arg Glu Ala Ser Asn Cys 35 40 45 Phe Ala Ile Arg His Phe Glu AsnLys Phe Ala Val Glu Thr Leu Ile 50 55 60 Cys Ser 65 15 180 PRT Homosapiens 15 Gly Gly Glu Glu Pro Ala Glu Glu Asp Ser Glu Asp Trp Cys ValPro 1 5 10 15 Cys Ser Asp Glu Glu Val Glu Leu Pro Ala Asp Gly Gln ProTrp Met 20 25 30 Pro Pro Pro Ser Glu Ile Gln Arg Leu Tyr Glu Leu Leu AlaAla His 35 40 45 Gly Thr Leu Glu Leu Gln Ala Glu Ile Leu Pro Arg Arg ProPro Thr 50 55 60 Pro Glu Ala Gln Ser Glu Glu Glu Arg Ser Asp Glu Glu ProGlu Ala 65 70 75 80 Lys Glu Glu Glu Glu Glu Lys Pro His Met Pro Thr GluPhe Asp Phe 85 90 95 Asp Asp Glu Pro Val Thr Pro Lys Asp Ser Leu Ile AspArg Arg Arg 100 105 110 Thr Pro Gly Ser Ser Ala Arg Ser Gln Lys Arg GluAla Arg Leu Asp 115 120 125 Lys Val Leu Ser Asp Met Lys Arg His Lys LysLeu Glu Glu Gln Ile 130 135 140 Leu Arg Thr Gly Arg Asp Leu Phe Ser LeuAsp Ser Glu Asp Pro Ser 145 150 155 160 Pro Ala Ser Pro Pro Leu Arg SerSer Gly Ser Ser Leu Phe Pro Arg 165 170 175 Gln Arg Lys Tyr 180 16 19PRT Homo sapiens 16 Arg Thr Val Ser Ser Ile Asn Gly Val Gly Lys Thr GlyTyr Pro Tyr 1 5 10 15 Ala Lys Glu 17 19 PRT Homo sapiens 17 Asn Leu GlyLeu Leu Phe Ile Leu Ala Thr Ser Ser Leu Ala Val Tyr 1 5 10 15 Ser IleLeu 18 30 PRT Homo sapiens 18 Lys Ser Met Lys Asn Asn Pro Val Ile ValPhe Ala Thr Lys Gly Lys 1 5 10 15 Gln Cys Lys Met Thr Tyr Ser Ile SerSer Cys Ser Asn Tyr 20 25 30 19 163 PRT Homo sapiens 19 Asn Lys His AlaSer Phe Tyr Ser Ser Ser Asn Gln Lys Asn Lys Pro 1 5 10 15 Ser Phe HisArg Ser Cys His Gln Val Phe Pro His Ala Ser Asn Arg 20 25 30 Ile His AsnPro Ser Asn Ser Tyr Pro Leu Gln Gln Tyr Thr Leu Arg 35 40 45 Thr Met AsnHis Asn Gln Tyr Tyr Gln Ser Ile Leu Ile Ile Asn Asn 50 55 60 His Asn GlyTyr Ser Asn Lys Thr Arg Asn Ser Pro Leu Ser Leu Leu 65 70 75 80 Ser ProArg Gly Tyr Pro Arg His Pro Ser Asp Ile Arg Pro Ala Ser 85 90 95 Ser HisMet Thr Lys Thr Ser Pro His Leu Asn His Ile Pro Asn Leu 100 105 110 SerLeu Thr Lys Arg Lys Pro Ser Pro His Ser Leu Asn Leu Ile His 115 120 125His Ser Arg Gln Leu Arg Trp Ile Lys Pro Asn Pro Ala Thr Gln Asn 130 135140 Leu Ser Ile Leu Leu Asn Tyr Pro Tyr Arg Met Asn Asn Ser Ser Ser 145150 155 160 Thr Val Gln 20 105 PRT Homo sapiens 20 Lys Asn Ala Leu AlaHis Phe Leu Pro Gln Gly Thr Pro Thr Pro Leu 1 5 10 15 Ile Pro Ile LeuVal Ile Ile Glu Thr Ile Ser Leu Leu Ile Gln Pro 20 25 30 Ile Ala Leu AlaVal Arg Leu Thr Ala Asn Ile Thr Ala Gly His Leu 35 40 45 Leu Met His LeuIle Gly Ser Ala Thr Leu Ala Ile Ser Thr Ile Asn 50 55 60 Leu Pro Ser ThrLeu Ile Ile Phe Thr Ile Leu Ile Leu Leu Thr Ile 65 70 75 80 Leu Glu IleAla Val Ala Leu Ile Gln Ala Tyr Val Phe Thr Leu Leu 85 90 95 Val Ser LeuTyr Leu His Asp Asn Thr 100 105 21 117 PRT Homo sapiens 21 Ala Arg ProIle Leu Arg Ile Ile Ser Leu Leu Arg Asn Leu Lys His 1 5 10 15 Arg HisTyr Pro Pro Ala Cys Asn Tyr Ser Asn Ser Leu His Arg Leu 20 25 30 Cys ProPro Val Arg Pro Asn Ile Ile Leu Arg Gly His Ser Asn Tyr 35 40 45 Lys LeuThr Ile Arg His Pro Ile His Trp Asp Arg Pro Ser Ser Met 50 55 60 Asn LeuArg Arg Leu Leu Ser Arg Gln Ser His Pro His Thr Ile Leu 65 70 75 80 TyrLeu Ser Leu His Leu Ala Leu His Tyr Cys Ser Pro Ser Ser Thr 85 90 95 ProPro Pro Ile Leu Ala Arg Asn Gly Ile Lys Gln Pro Pro Arg Asn 100 105 110His Leu Pro Phe Arg 115 22 24 PRT Homo sapiens 22 Glu Pro Ser Gly SerCys Ala Asn Arg His Gln Tyr Gln Leu Cys Glu 1 5 10 15 Leu Gly Cys GluCys Thr Arg Ala 20 23 520 PRT Homo sapiens 23 Pro Thr Asp Ser Thr MetLeu Lys Lys Phe Asp Lys Lys Asp Glu Glu 1 5 10 15 Ser Gly Gly Gly SerAsn Pro Phe Gln His Leu Glu Lys Ser Ala Val 20 25 30 Leu Gln Glu Ala ArgVal Phe Asn Glu Thr Pro Ile Asn Pro Arg Lys 35 40 45 Cys Ala His Ile LeuThr Lys Ile Leu Tyr Leu Ile Asn Gln Gly Glu 50 55 60 His Leu Gly Thr ThrGlu Ala Thr Glu Ala Phe Phe Ala Met Thr Lys 65 70 75 80 Leu Phe Gln SerAsn Asp Pro Thr Leu Arg Arg Met Cys Tyr Leu Thr 85 90 95 Ile Lys Glu MetSer Cys Ile Ala Glu Asp Val Ile Ile Val Thr Ser 100 105 110 Ser Leu ThrLys Asp Met Thr Gly Lys Glu Asp Asn Tyr Arg Gly Pro 115 120 125 Ala ValArg Ala Leu Cys Gln Ile Thr Asp Ser Thr Met Leu Gln Ala 130 135 140 IleGlu Arg Tyr Met Lys Gln Ala Ile Val Asp Lys Val Pro Ser Val 145 150 155160 Ser Ser Ser Ala Leu Val Ser Ser Leu His Leu Leu Lys Cys Ser Phe 165170 175 Asp Val Val Lys Arg Trp Val Asn Glu Ala Gln Glu Ala Ala Ser Ser180 185 190 Asp Asn Ile Met Val Gln Tyr His Ala Leu Gly Leu Leu Tyr HisVal 195 200 205 Arg Lys Asn Asp Arg Leu Ala Val Asn Lys Met Ile Ser LysVal Thr 210 215 220 Arg His Gly Leu Lys Ser Pro Phe Ala Tyr Cys Met MetIle Arg Val 225 230 235 240 Ala Ser Lys Gln Leu Glu Glu Glu Asp Gly SerArg Asp Ser Pro Leu 245 250 255 Phe Asp Phe Ile Glu Ser Cys Leu Arg AsnLys His Glu Met Val Val 260 265 270 Tyr Glu Ala Ala Ser Ala Ile Val AsnLeu Pro Gly Cys Ser Ala Lys 275 280 285 Glu Leu Ala Pro Ala Val Ser ValLeu Gln Leu Phe Cys Ser Ser Pro 290 295 300 Lys Ala Ala Leu Arg Tyr AlaAla Val Arg Thr Leu Asn Lys Val Ala 305 310 315 320 Met Lys His Pro SerAla Val Thr Ala Cys Asn Leu Asp Leu Glu Asn 325 330 335 Leu Val Thr AspSer Asn Arg Ser Ile Ala Thr Leu Ala Ile Thr Thr 340 345 350 Leu Leu LysThr Gly Ser Glu Ser Ser Ile Asp Arg Leu Met Lys Gln 355 360 365 Ile SerSer Phe Met Ser Glu Ile Ser Asp Glu Phe Lys Val Val Val 370 375 380 ValGln Ala Ile Ser Ala Leu Cys Gln Lys Tyr Pro Arg Lys His Ala 385 390 395400 Val Leu Met Asn Phe Leu Phe Thr Met Leu Arg Glu Glu Gly Gly Phe 405410 415 Glu Tyr Lys Arg Ala Ile Val Asp Cys Ile Ile Ser Ile Ile Glu Glu420 425 430 Asn Ser Glu Ser Lys Glu Thr Gly Leu Ser His Leu Cys Glu PheIle 435 440 445 Glu Asp Cys Glu Phe Thr Val Leu Ala Thr Arg Ile Leu HisLeu Leu 450 455 460 Gly Gln Glu Gly Pro Lys Thr Thr Asn Pro Ser Lys TyrIle Arg Phe 465 470 475 480 Ile Tyr Asn Arg Val Val Leu Glu His Glu GluVal Arg Ala Gly Ala 485 490 495 Val Ser Ala Leu Ala Lys Phe Gly Ala GlnAsn Glu Glu Met Leu Pro 500 505 510 Ser Ile Leu Val Leu Leu Lys Arg 515520 24 21 PRT Homo sapiens 24 Pro Gln Thr His Ser Thr Leu Leu Pro AspAsn Leu Ser Gln Thr Ile 1 5 10 15 Tyr Pro Asn Lys Val 20 25 46 PRT Homosapiens 25 His Asn Lys Gln Thr Trp Ser Ile Leu Gly Leu Leu His Ala LeuPro 1 5 10 15 Glu Pro Val Pro Ala Ala Gly Gly Ser Pro Gly Glu Arg LeuTrp Pro 20 25 30 Leu Gln Leu Leu Arg Glu Cys Ala Arg Gln Thr Arg Gln Ile35 40 45 26 34 PRT Homo sapiens 26 Asn Thr Lys Ile Cys Gln Ala Trp TrpArg Thr Pro Ile Ile Pro Ala 1 5 10 15 Thr Gly Glu Ala Glu Ala Gly GluPro Leu Glu Pro Arg Arg Gln Arg 20 25 30 Leu Gln 27 180 PRT Homo sapiens27 Gly Leu Leu Lys Val Val Phe Val Val Phe Ala Ser Leu Cys Ala Trp 1 510 15 Tyr Ser Gly Tyr Leu Leu Ala Glu Leu Ile Pro Asp Ala Pro Leu Ser 2025 30 Ser Ala Ala Tyr Ser Ile Arg Ser Ile Gly Glu Arg Pro Val Leu Lys 3540 45 Gly Glu Cys Arg Ala Trp Gly Arg Arg Leu Pro Ser Trp Leu Val Cys 5055 60 Arg Gly Arg Ser Gly Gly Phe Cys Pro Ser Trp Arg Leu Gly Gly Pro 6570 75 80 Asp Gly Phe Ile Ser Gly Arg Arg Arg Arg Glu Ala Phe Cys Ser Tyr85 90 95 Ser Phe Arg Ser His Ala Ile Pro Arg His Val Pro Leu Leu Thr Pro100 105 110 Cys Glu Thr Arg Thr Val Trp Leu Leu Val Thr Ser Thr Asn LeuGln 115 120 125 Pro Asp Leu Glu Leu Cys Ser Phe Arg Pro Asp Leu Gly PheLeu Pro 130 135 140 Trp Glu Ala Gln Glu Gly Pro Gly Ser Glu Ile Pro AsnHis Phe Lys 145 150 155 160 Val Ser Val Gly Leu Lys Ser Cys Trp Lys IlePhe Cys Lys Val Leu 165 170 175 Gly Ser Lys Ser 180 28 34 PRT Homosapiens 28 Lys Lys Lys Ser Thr Trp Val Gln Trp Leu Thr Pro Val Ile LeuAla 1 5 10 15 Phe Gly Glu Thr Lys Val Gly Gly Ser Leu Glu Pro Gly ArgSer Arg 20 25 30 Leu Gln 29 7 PRT Homo sapiens 29 Arg Ser Cys His AspLeu Asn 1 5 30 237 PRT Homo sapiens 30 Tyr Ala Ala Glu Ile Cys Ile AlaLeu Asn Phe Leu His Glu Arg Gly 1 5 10 15 Ile Ile Tyr Arg Asp Leu LysLeu Asp Asn Val Leu Leu Asp Ala Asp 20 25 30 Gly His Ile Lys Leu Thr AspTyr Gly Met Cys Lys Glu Gly Leu Gly 35 40 45 Pro Gly Asp Thr Thr Ser ThrPhe Cys Gly Thr Pro Asn Tyr Ile Ala 50 55 60 Pro Glu Ile Leu Arg Gly GluGlu Tyr Gly Phe Ser Val Asp Trp Trp 65 70 75 80 Ala Leu Gly Val Leu MetPhe Glu Met Met Ala Gly Arg Ser Pro Phe 85 90 95 Asp Ile Ile Thr Asp AsnPro Asp Met Asn Thr Glu Asp Tyr Leu Phe 100 105 110 Gln Val Ile Leu GluLys Pro Ile Arg Ile Pro Arg Phe Leu Ser Val 115 120 125 Lys Ala Ser HisVal Leu Lys Gly Phe Leu Asn Lys Asp Pro Lys Glu 130 135 140 Arg Leu GlyCys Arg Pro Gln Thr Gly Phe Ser Asp Ile Lys Ser His 145 150 155 160 AlaPhe Phe Arg Ser Ile Asp Trp Asp Leu Leu Glu Lys Lys Gln Ala 165 170 175Leu Pro Pro Phe Gln Pro Gln Ile Thr Asp Asp Tyr Gly Leu Asp Asn 180 185190 Phe Asp Thr Gln Phe Thr Ser Glu Pro Val Gln Leu Thr Pro Asp Asp 195200 205 Glu Asp Ala Ile Lys Arg Ile Asp Gln Ser Glu Phe Glu Gly Phe Glu210 215 220 Tyr Ile Asn Pro Leu Leu Leu Ser Thr Glu Glu Ser Val 225 230235 31 54 PRT Homo sapiens 31 Phe Trp Ile Leu Pro Ser Ile Ser Arg ProLeu Arg Ala Arg His Phe 1 5 10 15 Arg Ala Gly Gln Asn Leu Cys Arg SerArg Cys Gly Phe Ala Gly Asp 20 25 30 Gln Gly Gly Asp Glu Lys Gly Arg GlyGly Ala Pro Ala Gln Glu Arg 35 40 45 Gly Cys Gly Ala Pro Gly 50 32 26PRT Homo sapiens 32 Lys Lys Lys Lys Tyr Leu Gly Thr Val Ala His Thr CysAsn Pro Ser 1 5 10 15 Ile Trp Gly Asp Gln Gly Gly Arg Ile Thr 20 25 331145 DNA Homo sapiens 33 gttgtagata tgcggcgtta tttctgaggg ctctgttctgttccattgat ctatatctct 60 gtttcggtac cagtaccatg ctgttactgt agccttgtagtatagtttga aatcaggtag 120 cgtgatgcct ccagctttgt tctttcggct taggattgacttggcaatgc gggctctttt 180 ttggttccat atgaacttta aagtagtttt ttccaattctgtgaagaaag tgattggtag 240 cttgatgggg atggcattga atctataaat taccttaggcagtatggcca ttttcacgat 300 attgattctt cctacccatg agcatggaat gttcttccatttctttgtat cctcttttat 360 ttcattgagc agtggtttgt agttctcctt gatgaggtccttcacaaaaa ctaacaaaca 420 gaaaggacgt gacagcccag agtttttaaa gctgacacttgagttgggtt tttctctttc 480 tttaccactg taaaaaaata atgaatgaga attgggtgctgggttttgtt ttgttgtttt 540 agtggtggca gggaagctgg ggatgcgggt gattaggctgagtattacag aggtagcagg 600 agtcccatag caccaagttc catcttcttg ttacccattccagaggcccc aaacctacct 660 agaggctttg ggatcaggag gcttgagctt ggattctggctctgccacct ccttacctag 720 aacccctttg taagtcactg agtttgtgag tctcaacgtcgtcattacaa aagtacagaa 780 acaccaacct tctctttcat ggtgggtagt gtggtagtactggactctgg atgagttata 840 ctttaaaagc tatcaactgt atatcatgta atcctgcccctacaggactg attcactaac 900 aagataactg tttaaaagaa agaaaaagag agcggggcgcagtggctcac acctgtaatc 960 ccagcacttt gggaggctga ggcgggtgga tcatttgaggtcaggagttt gaggccagcc 1020 tggccaacat agtgaaaccc tgtctctacc agaaatacaaaaaaattagc cgggtgtggt 1080 agtgcatgcc tataatccca gctacttggg aggctgaggcaggagaatca cttgaacccg 1140 ggagg 1145 34 929 DNA Homo sapiens 34aacactgcca gtatgttact gagaaggtcc tggctgctgt ctacaaggcc ctgaatgacc 60atcatgttta cctggagggc accctgctaa agcccaacat ggtgactgct ggacatgcct 120gcaccaagaa gtatactcca gaacaagtag ctatggccac cgtaacagct ctccaccgta 180ctgttcctgc agctgttcct ggcatctgct ttttgtctgg tggcatgagt gaagaggatg 240ccactctcaa cctcaatgct atcaaccttt gccctctacc aaagccctgg aaactaagtt 300tctcttatgg acgggccctg caggccagtg cactggctgc ctggggtggc aaggctgcaa 360acaaggaggc aacccaggag gcttttatga agcgggccat ggctaactgc caggcggcca 420aaggacagta tgttcacacg ggttcttctg gggctgcttc cacccagtcg ctcttcacag 480cctgctatac ctactagggt ccaatgcccg ccagcctagc tccagtgctt ctagtaggag 540ggctgaaagg gagcaacttt tcctctaatc ctggaaattc gacacaatta gatttgaact 600gctggaaata caacacatgt taaatcttaa gtacaagggg gaaaaaataa atcagttatt 660gaaacataaa aatgaatacc aaggacctga tcaaatttca cacagcagtt tccttgcaac 720actttcagct ccccatgctc cagaataccc acccaagaaa ataataggct ttaaaacaat 780atcggctcct catccaaaga acaactgctg attgaaacac ctcattagct gagtgtagag 840aagtgcatct tatgaaacag tcttagcagt ggtaggttgg gaaggagata gctgcaacca 900aaaaagaaat aaatattcta taaaccttc 929 35 479 DNA Homo sapiens 35cataatcata ggtagctagg ttataaacta tttaaagaca agatcacgtg ataagcttat 60aatcttctca taattcccct tacttagcat tgtgttagac atactaatag gtgcacagtg 120aaatacttat tgttgattgt ttaaaaataa agttttagaa aaccttttca aaagtcagag 180tttaggccag gggcacaggc tgacacctat aatcccagca ctttgggagg ccagggcggg 240cagatcactt gggtcaagag ttcaaggcca gcctggccaa catggcaaaa ccccatctct 300actaaataaa atacaaaaat tatccaggca tggtggtgca tgcctgtaat cccagctact 360tggaggctga ggcatgagaa ttgcttgaac ctgggaggca gaggttgcag tgagctgaga 420tcgccccact gcaatccagc ctgggagaca taattcaaat ctattttggt cttatatct 479 36253 DNA Homo sapiens 36 ctctcttttt ctaatgtaaa tgttgtgtac aatagttttatttgattaag cttcaggact 60 gttttgtaaa gcgaggtggg accgatgtgg cacacgccagctgcggtttc ccggagcgtg 120 gagaggcagt gctgctgctc ccgcccgagg ctcatgacaactcaataaag cactgctttt 180 attttttgca gtcttcaatt tgagaaaggt gagaaataatgttttccaat aaatgagatt 240 cataccatta aaa 253 37 1884 DNA Homo sapiens 37tctggtggcc cttccggcca cccctttaac cccagctttc cctccccctt ctttcgatca 60gagatcggcg gagaccctcg aagtgcgcaa acttgacact caccctgacc ggactggggt 120tttaaggggt gtggcaggag gttttggact cgatgagttt ccaccgaaat gtcggagaag 180tcaggccaga gcacaaaagc aaaggatggg aaaaagtatg caacactcag tttatttaat 240acttacaagg ggaaatcatt agaaacacag aaaaccacag ctcgacatgg attacagagt 300cttggaaaag tcggtatttc acggcgtatg cctccacctg ctaacctccc aagtcttaaa 360gcagaaaaca aaggcaatga tcctaatgta aacattgtac ctaaagatgg cacagggtgg 420gcatcaaaac aagagcaaca tgaagaagaa aaaacaccag aagtgccacc agcacagcca 480aaacctgggg ttgcagctcc cccagaagta gcacctgctc ccaaatcatg ggccagtaac 540aagcaaggtg ggcaaggaga tggaatccaa gtgaatagtc agtttcagca agaatttccc 600agcctgcagg cagctgggga tcaggaaaaa aaagaaaagg aaacaaatga tgacaactat 660ggacctggac ccagtttacg tccaccaaat gttgcttgtt ggagagatgg tggtaaggct 720gctggctcac cttcgtcatc tgatcaagat gaaaagctcc ctggccagga tgaaagcaca 780gctggaacat cagagcaaaa tgatatcctc aaagtggtgg aaaagaggat agcttgtggt 840cctccacagg ctaaactgaa tggacagcag actgctctcg cttcccagta tagagctatg 900atgcctcctt atatgttcca acagtatccg aggatgacat atcctcctct acatggtccc 960atgagattcc caccttcttt atctgaaaca aacaaaggcc ttcgaggaag aggcccacct 1020ccttcatggg cctctgagcc tgaacgccca tccattctta gtgcatcaga actgaaggag 1080cttgataaat ttgataacct agatgctgaa gctgatgaag gttgggcagg tgctcagatg 1140gaagtagatt atacagagca actgaatttc agtgatgatg atgaacaagg aagtaacagt 1200cctaaagaga ataacagtga ggatcaaggt tcaaaagcct ctgaaaacaa cgaaaacaaa 1260aaagaaacag atgaagtttc caacactaaa tcatcttccc aaatacctgc ccaaccatca 1320gtagcaaaag ttccctatgg gaaaggacct tcatttaatc aggaacgtgg aacatcttca 1380catctgccac cacctccaaa gttgcttgca cagcagcatc cacctccaga tcgacaggca 1440gtacctggaa gaccaggccc ctttccctcc aagcagcaag tagctgatga agatgaaata 1500tggaagcaaa gacgaagaca acaatcagaa atttctgcag cagtagaacg tgctcgtaaa 1560cggcgtgaag aggaagagcg aagaatggaa gaacaaagga aggcagcttg tgcggagaaa 1620ctgaaacgat tggatgagaa gcttggcatc ctggaaaaac aaccatctcc agaggaaatt 1680agggaaaggg agcgagaaaa agaacgggag cgtgagaaag aacttgaaaa agaacaagaa 1740caggagcgag agaaggagag ggaaaaagac agagagagac agcaggaaaa ggagaaagag 1800ctggagaagg agcaggaaaa acaaagagaa atggagaaag aaagaaagca agaaaaagaa 1860aaagaactag aacggcagaa aaaa 1884 38 1004 DNA Homo sapiens 38 cgcaaaattaaccccctaat aaaattaatt aaccactcat tcatcgacct ccccacccca 60 tccaacatctccgcatgatg aaacttcggc tcactccttg gcgcctgcct gatcctccaa 120 atcaccacaggactattcct agccatgcac tactcaccag acgcctcaac cgccttttca 180 tcaatcgcccacatcactcg agacgtaaat tatggctgaa tcatccgcta ccttcacgcc 240 aatggcgcctcaatattctt tatctgcctc ttcctacaca tcgggcgagg cctatattac 300 ggatcatttctctactcaga aacctgaaac atcggcatta tcctcctgct tgcaactata 360 gcaacagccttcataggcta tgtcctcccg tgaggccaaa tatcattctg aggggccaca 420 gtaattacaaacttactatc cgccatccca tacattggga cagacctagt tcaatgaatc 480 tgaagaggctactcagtaga cagtcccacc ctcacacgat tctttacctt tcacttcatc 540 ttgcccttcattattgcagc cctagcagca ctccacctcc tattcttgca cgaaacggga 600 tcaaacaaccccctaggaat cacctcccat tccgataaaa tcaccttcca cccttactac 660 acaatcaaagacgccctcgg cttacttctc ttccttctct ccttaatgac attaacacta 720 ttctcaccagacctcctagg cgacccagac aattataccc tagccaaccc cttaaacacc 780 cctccccacatcaagcccga atgatatttc ctattcgcct acacaattct ccgatccgtc 840 cctaacaaactaggaggcgt ccttgcccta ttactatcca tcctcatcct agcaataatc 900 cccatcctccatatatccaa acaacaaagc ataatatttc gcccactaag ccaatcactt 960 tattgactcctagccgcaga cctcctcatt ctaacctgaa tcgg 1004 39 1129 DNA Homo sapiens 39aaagtgtgag agggtccgta gttgggtcaa ctttgactcc tctcgcctgc ccggatcctt 60aagggcctcc tcgtcctccc ggtctccggt cgctgccggg tctgtgcgcc ggtccgcgcc 120cgccctcgct ctgccatggg cgcttccagc tcctccgcgc tggcccgcct cggcctccca 180gcccggccct ggcccaggtg gctcggggtc gccgcgctag gactggccgc cgtggccctg 240gggactgtcg cctggcgccg cgcatggccc aggcggcgcc ggcggctgca gcaggtgggc 300accgtggcga agctctggat ctacccggtg aaatcctgca aaggggtgcc ggtgagcgag 360gctgagtgca cggccatggg gctgcgcagc ggcaacctgc gggacaggtt ttggctggtg 420attaaggaag atggacacat ggtcactgcc cgacaggagc ctcgcctcgt gctcatctcc 480atcatttatg agaataactg cctgatcttc agggctccag acatggacca gctggttttg 540cctagcaagc agccttcctc aaacaaactc cacaactgca ggatatttgg ccttgacatt 600aaaggcagag actgtggcaa tgaggcagct aagtggttca ccaacttctt gaaaactgaa 660gcgtatagat tggttcaatt tgagacaaac atgaagggaa gaacatcaag aaaacttctc 720cccactcttg atcagaattt ccaggtggcc tacccagact actgcccgct cctgatcatg 780acagatgcct ccctggtaga tttgaatacc aggatggaga agaaaatgaa aatggagaat 840ttcaggccaa atattgtggt gaccggctgt gatgcttttg aggaggcttc agcaaccagg 900agggattgac tgagatctta acaacagcag caacgataca tcagcaaatc cttattatcc 960agccttcaac tatctttacc ctggaaaaca atctcgattt ttgacttttc aaagttgtgt 1020atgctccagg ttaatgcaag gaaagtatta gaggggggaa tatgaaagta tatatataaa 1080ttttaggtac tgaaggcttt aaaaataatt aagatcatca aaaatgcta 1129 40 821 DNAHomo sapiens 40 gtcaactatg ctggctggac tggctgcctt gttcctggcc taggacttagcttcataact 60 atcacctgca ccgactaggc tgaggtgctg gtacttgccc caacccctacttttgtattt 120 atatgtgtgt gtgtgtgtgc gtgcgtgcgt gcgtgcgtgt atgtttggtctggaccagct 180 tctgccagcc cctggccttt actttcttcc ttgcctatgc agggcaaacaaaatgtgaaa 240 ttctgccctc agctgagctg agtaagggct cctgggggtt ggctggagatgggtgtggca 300 tctgtccagg cctggaaccg tctcaagaca gtgctggcaa agctgcagtattgagatgct 360 aaggagctga tgccacctct ttgtcttccc ctaaaggaga acatggggataacatgggtg 420 tgtgcccaca acactctagg tgcagagccc ctgtggcaaa gtattacagggtgtgggtgg 480 ggattaccct gaatcgggga ttttaatgat ggaagcaggc agagcctggtgggtgattct 540 gtcaacagaa aattgcaatc atgcaggggc tgggagggtt aggatgaaaaaactggggcc 600 attggaggcc cactgtaggt gggagggagc tgattttggg gtggggggtgggactagagg 660 gcaatactga aggggttaaa caggtttttg ctcctcaaga atttgtttgcctgggcccag 720 gattggaggg cttcacacca ataccctgtg tatacaagaa tcagatttataatacttccc 780 cttttttgtt acgtatgaac actataaacc aaattatttt g 821 41 544DNA Homo sapiens 41 tcatttggct cactatgaaa agcctttaat taatctcttcaaacaagtta tttccttaat 60 ccacaagcag tggttacact tgctttgcat tcttgtctggttcctaactc tagagccctt 120 ctccctggct tagccagtaa gctgagcccc tggctgcgttcagccggccc gcctgagaga 180 cactagggga aatagctttt gtgggcaagc agggtggccggtggtgctca gcagtctttc 240 cagtggctgt gtccctcctc caaatgtgga caggccatgacagagtctta gcccaagtcc 300 cacagatccc caaaagttct gttgattgct tcaggggatcagtgaaaatt agggaatttt 360 gtgtgttgct atatacattt tttctgggga gatgagcttctcattgagat ctgtgactca 420 gaatcgacta agccaccata agtctggatt tctccccagctcccaaggcc cttttgggtc 480 cagaagacct gcatatgggc tgttgactca tgcaaatgaggtatctgaac tgcagcttca 540 gtat 544 42 1067 DNA Homo sapiens 42cctgtcagtc gccatcatac ctcgagtgag gcccagctag ataatgactt gtccaagatg 60gcacacgtgg aaagttgatc tgcaccagaa cccggatgac tgtcaccttg aagcgtcctg 120ttctccttct gtgctgtccc aggaagtgtc tggcgggcgt gggcagcaca gctctacact 180gtacgattca ctagggcatc ctgcgagcct cactagcctt ctggttcatg cctttgacaa 240gcatttttgt gccccctctg cttactgtga cagtcgatga tgaatcttgc gttgccattt 300tctgctgtgg gtaactgcgt gcagtgtctt gccttgcttt ctcttcttac tgtcccacag 360cttggtttca tgttacaaac agaaaagctc gaggctccca ccccgccaca tcccaacttc 420atttccccct cactgtagcc catttccacc ccaccacaaa gttgccacag gttttctttg 480tatagaatat ttattttgaa gctctatttt aatagtattt attttagaaa gtctactatt 540gtaagagttc ttctgtttgt gaagaaaaaa acaagttaaa aactgaatgt actgatttag 600aaaatatata taaatatata ttgttaaata tactttgatt gcgccactgc actccagcct 660tggcgaccag actaagacgc tgtctcaaaa aaaaacaaaa acgacaaaaa aaaaacaaaa 720cagaaaaaat aaactaaggc aatgacagtc cctggcaaat gctgggaggg aggcagcagt 780ggtcagggaa ggtaaccctg aagcaggact tgtaaagcaa ataagattgg gaggccaagg 840tgggtggatc acgaggtcag gagttcgaga ccagcctggc caacatagtg aaaccccgtc 900tttactaaaa atacaaaaaa attagccagg tgtggtggtg ggtgcctgta gtcccagcta 960cttgggaggc tgaggcagga gaatctcgaa cccaggaggc ggaggttaca gtcagctgag 1020accgcaccat tgcactccag cctgggtgac agagcaagat tccgtct 1067 43 1560 DNAHomo sapiens 43 ttaagggaaa aaatgactgg ctatattcag tcagatatgg caaaaagtctcaaggtgtta 60 atgtgaatga ttaaggtctt ggggggggtg tcccctatca gactacaggtgtttagaggc 120 acagaaaaag gtgcagttgg gttcttaatg tgaaatgatg agaagcacaactccagtgtg 180 tctctttgtg tagaatgtca gcagacaccc cctgctagat gtgctggatcatgggaaagc 240 atttccattt gttactagat tgttcagaag ttttaattta tgatgggtgtggtggctcat 300 gcctgtagtc ccagcactgt gggaggctga ggcaggagga tcatctgaggccaagagttc 360 aagatcagcc tgggcaacat agtgataccc tatctcttaa aaaagaagaagtttttaaat 420 ttgaaataat aataggtact ggatttatgc aaatgtcttt tctgcgtcttttgagatgag 480 tatcaggttt ttttttttcc ttttatcatc tgatgatgaa cttaatgtttccatttgtat 540 taatggaata ctaagtccct ctgtgatttc tgaaccaagc tattcctaggcctgagtttt 600 attttgttga cacagaaata aattagaagg ccaagcgtgg tggcatgtgcctgtagtcct 660 agttgctgag gtaagaggat tgcttgagcc caggagttca aggctgcagcaagctttgat 720 tgcgccactg cactccagcc ttggcgacag actaagacgc tgtctcaaaaaaaaacaaaa 780 acgacaaaaa aaaaacaaaa cagaaaaaat aaactaaggc aatgacagtccctggcaaat 840 gctgggaggg aggcagcagt ggtcagggaa ggtaaccctg aagcaggacttgtaaagcaa 900 ataagattgg gaggccaagg tgggtggatc acgaggtcag gagttcgagaccagcctggc 960 caacatagtg aaaccccgtc tttactaaaa atacaaaaaa attagccaggtgtggtggtg 1020 ggtgcctgta gtcccagcta cttgggaggc tgaggcagga gaatctcgaacccaggaggc 1080 ggaggttaca gtcagctgag accgcaccat tgcactccag cctgggtgacagagcaagat 1140 tccgtctcaa aaaaaaaaaa aaaaaaaaaa ccaagaagaa aaggaatgaattagaacttc 1200 ttctgcttgg acttaagggc atcatcaggc aggttttggg taggatagcaggggaggcag 1260 agacatagtc ggggtcagtg gtcatgagtg tggctttgag cccaaaaacttggtttctgt 1320 tccctacttt gccactcagt agtgcatgac tttggccaaa tttcttaaattcatgaagca 1380 agtttccggg tgaatgaaat ggggataaaa atagtgttca aacctatccgttggtttgtg 1440 tgaaactgaa atgaatagta tcgtgcaggt acttgtgagc aaggggagctgctgtttcct 1500 gtccctttat gatgggaaat atctagacaa gttcccaacc ctctgcactgcaggctgcat 1560 44 397 DNA Homo sapiens 44 agagtcacta tccggaaatctaagaatatc ctctttgtca tcacaaaacc agatgtctac 60 aagagccctg cttcagatacttacatagtt tttggggaag ccaagatcga agatttatcc 120 cagcaagcac aactagcagctgctgagaaa ttcaaagttc aaggtgaagc tgtctcaaac 180 attcaagaaa acacacagactccaactgta caagaggaga gtgaagagga agaggtcgat 240 gaaacaggtg tagaagttaaggacatagaa ttggtcatgt cacaagcaaa tgtgtcgaga 300 gcaaaggcag tccgagccctgaagaacaac agtaatgata ttgtaaatgc gattatggaa 360 ttaacaatgt aaccatatggaagcaacttt ttttggt 397 45 931 DNA Homo sapiens 45 gcctagccat gtgatttcacttccactcca taacgctcct catactaggc ctactaacca 60 acacactaac catataccaatgatggcgcg atgtaacacg agaaagcaca taccaaggcc 120 accacacacc acctgtccaaaaaggccttc gatacgggat aatcctattt attacctcag 180 aagttttttt cttcgcaggatttttctgag ccttttacca ctccagccta gcccctaccc 240 cccaattagg agggcactggcccccaacag gcatcacccc gctaaatccc ctagaagtcc 300 cactcctaaa cacatccgtattactcgcat caggagtatc aatcacctga gctcaccata 360 gtctaataga aaacaaccgaaaccaaataa ttcaagcact gcttattaca attttactgg 420 gtctctattt taccctcctacaagcctcag agtacttcga gtctcccttc accatttccg 480 acggcatcta cggctcaacattttttgtag ccacaggctt ccacggactt cacgtcatta 540 ttggctcaac tttcctcactatctgcttca tccgccaact aatatttcac tttacatcca 600 aacatcactt tggcttcgaagccgccgcct gatactggca ttttgtagat gtggtttgac 660 tatttctgta tgtctccatctattgatgag ggtcttactc ttttagtata aatagtaccg 720 ttaacttcca attaactagttttgacaaca ttcaaaaaag agtaataaac ttcgccttaa 780 ttttaataat caacaccctcctagccttac tactaataat tattacattt tgactaccac 840 aactcaacgg ctacatagaaaaatccaccc cttacgagtg cggcttcgac cctatatccc 900 ccgcccgcgt ccctttctccataaaattct t 931 46 968 DNA Homo sapiens 46 gagcacatgg ttattactgatcgcattgaa aacattgatc acctgggttt ctttatttat 60 cgactgtgtc atgacaaggaaacttacaaa ctgcaacgca gagaaactat taaaggtatt 120 cagaaacgtg aagccagcaattgtttcgca attcggcatt ttgaaaacaa atttgccgtg 180 gaaactttaa tttgttcttgaacagtcaag aaaaacatta ttgaggaaaa ttaatatcac 240 agcataaccc caccctttacattttgtgca gtgattattt tttaaagtct tctttcatgt 300 aagtagcaaa cagggctttactatcttttc atctcattaa ttcaattaaa accattacct 360 taaaattttt ttctttcgaagtgtggtgtc ttttatattt gaattagtaa ctgtatgaag 420 tcatagataa tagtacatgtcaccttaggt agtaggaaga attacaattt ctttaaatca 480 tttatctgga tttttatgttttattagcat tttcaagaag acggattatc tagagaataa 540 tcatatatat gcatacgtaaaaatggacca cagtgactta tttgtagttg ttagttgccc 600 tgctacctag tttgttagtgcatttgagca cacattttaa ttttcctcta attaaaatgt 660 gcagtatttt cagtgtcaaatatatttaac tatttagaga atgatttcca cctttatgtt 720 ttaatatcct aggcatctgctgtaataata ttttagaaaa tgtttggaat ttaagaaata 780 acttgtgtta ctaatttgtataacccatat ctgtgcaatg gaatataaat atcacaaagt 840 tgtttaacta gactgcgtgttgtttttccc gtataataaa accaaagaat agtttggttc 900 ttcaaatctt aagagaatccacataaaaga agaaactatt ttttaaaaat tcacttctat 960 atatacaa 968 47 904 DNAHomo sapiens 47 ctgggggaga agagcctgcc gaggaggact ccgaggactg gtgcgtgccctgcagcgacg 60 aggaggtgga gctgcctgcg gatgggcagc cctggatgcc cccgccctccgaaatccagc 120 ggctctatga actgctggct gcccacggta ctctggagct gcaagccgagatcctgcccc 180 gccggcctcc cacgccggag gcccagagcg aagaggagag atccgatgaggagccggagg 240 ccaaagaaga ggaagaggaa aaaccacaca tgcccacgga atttgattttgatgatgagc 300 cagtgacacc aaaggactcc ctgattgacc ggagacgcac cccaggaagctcagcccgga 360 gccagaaacg ggaggcccgc ctggacaagg tgctgtcgga catgaagagacacaagaagc 420 tggaggagca gatccttcgt accgggaggg acctcttcag cctggactcggaggacccca 480 gccccgccag ccccccactc cgatcctccg ggagtagtct cttccctcggcagcggaaat 540 actgattccc actgctcctg cctctagggt gcagtgtccg tacctgctggagcctgggcc 600 ctccttcccc agcccagaca ttgagaaact tgggaagaag agagaaacctcaagctccca 660 aacagcacgt tgcgggaaag aggaagagag agtgtgagtg tgtgtgtgtgttttttctat 720 tgaacacctg tagagtgtgt gtgtgtgttt tctattgaac acctatagagagagtgtgtg 780 tgttttctat tgaacatcta tatagagaga gtgtgtgagt gtgtgttttctattgaacac 840 ctattcagag acctggactg aattttctga gtctgaaata aaagatgcagagctatcatc 900 tctt 904 48 877 DNA Homo sapiens 48 aaaggacagt ctcttcaataaatggtgttg ggaaaactgg atatccatat gcaaaagaat 60 gaaattggac ctttatctcacaccattagc aaaaatcagc tcaaaatgaa ttaaagactt 120 aaacataaga cctgaaattgtaaaactact ggaagaaaac gggaaaagtt ctaccacatt 180 tgtctgcaaa atgtcatatatattcaacag aatactattc agccctagaa aagaaggaaa 240 tcctgtcatt tgtgacaatatgaatgaacc tggaagacat tatattaagc aaaataagcc 300 aggcatggaa agacaatattgcatgatctc acttatatgt ggaatctaaa aaaaattgga 360 cttacaaaag aaaaaaagagaatggtggtt actagatgcc ggggatggga atgcggatgg 420 ggaatgcaga gatgttgatcaaaaggtaca aagtttcagc tgggtgcagt ggctcacgcc 480 tgtaatccca gcactttgggaggccaaggc aggcagatca cttgaggtca ggagtttgag 540 accagcctgg ccaacatggtgaaaccctgt ccttgctaaa ataaaaaaat tagcccagtg 600 tggtggcatg cacctgtaatcccagctact caggaggtcg aggcggcaga attgcttgaa 660 cccaggaggt ggaggttgtagtgagctgag attgtgccac tgcactccag cctgggtgac 720 agagcaagac tctgtctcaaaaaaaaaaaa gtacgaagtt tcagttagac aggaaaagtg 780 agttttcagg acctattgaacagcatggtg accatagtta ataataatat atatttcaga 840 ttgctaaaag agcttattttaagtgttctt actacaa 877 49 793 DNA Homo sapiens 49 acgttgtagg cccctacgggctactacaac ccttcgctga cgccataaaa ctcttcacca 60 aagagcccct aaaacccgccacatctacca tcaccctcta catcaccgcc ccgaccttag 120 ctctcaccat cgctcttctactatgaaccc ccctccccat acccaacccc ctggtcaacc 180 tcaacctagg cctcctatttattctagcca cctctagcct agccgtttac tcaatcctct 240 gatcagggtg agcatcaaactcaaactacg ccctgatcgg cgcactgcga gcagtagccc 300 aaacaatctc atatgaagtcaccctagcca tcattctact atcaacatta ctaataagtg 360 gctcctttaa cctctccacccttatcacaa cacaagaaca cctctgatta ctcctgccat 420 catgaccctt ggccataatatgatttatct ccacactagc agagaccaac cgaaccccct 480 tcgaccttgc cgaaggggagtccgaactag tctcaggctt caacatcgaa tacgccgcag 540 gccccttcgc cctattcttcatagccgaat acacaaacat tattataata aacaccctca 600 ccactacaat cttcctaggaacaacatatg acgcactctc ccctgaactc tacacaacat 660 attttgtcac caagaccctacttctaacct ccctgttctt atgaattcga acagcatacc 720 cccgattccg ctacgaccaactcatacacc tcctatgaaa aaacttccta ccactcaccc 780 tagcattact tat 793 502086 DNA Homo sapiens 50 tattttcagt agagacgggg tttcaccatg ttagccaggatggtctcgat ctcctgacct 60 cgtgatccgc ccgcctcggc ctcccaaagt gctgggattacaggcatgag ccaccgggcc 120 cggatgtctt ggtttttctt gatatttgaa acagtaggtcaaaattggac atgtctagta 180 ggcggttgct ggtttagtaa taaagggtag tagctgtaattgcccagttt gaatagtgtt 240 catgttttta aggatttcac aaacaaatct tcatcgaataaatgttttta aaaaatgatt 300 actgattaga ttcaatataa attaaggaag gtcccatcatatctcttgca caaatcattg 360 ccttccagaa ttaagcacag catcgatatg tatgtcttacatgacatatt gctctgcaat 420 gtgatggaat caacagaaca agatgtttgg agtcagatggctaaactagg atttaaagtt 480 cacattattt actagttaac tgtggatgct aaggcaatattcttgaagtc tccatgtctt 540 gatttcctct tagtgaaatg ggaatcataa taataataattttacagggt tggtgtaaaa 600 acgaaatgac tgcctgcact caatacaata cctattatggttatgttagt tatattatga 660 ttaatgtatt agtatactat tgataaaatg aatatgaaattgctagtagc cagctcacaa 720 tattagtatg acaaaaatta tttcaatttg gtttaaatacaatacctgtg gtcaataaca 780 tcatcttttc ttaaaactta atgcactgag cactgtcaattacaatgaaa aaaaaatgca 840 agtaacccag ttggactatc cattaactga gaaaatataatatgaaaaag ccaccatgaa 900 ttcaggacca cttatgaatg tagctgtatt ttattgatagaatttatatg tgcttagaaa 960 aaacatagta cattttatgt tagatatatt tattcagattaccaaagaaa atgcttactg 1020 tacatatgga ttgatgggcc ctatccaata actttcatcactcctcaagg gcccttggca 1080 cacaggctgg gaatcactga tgttggttaa ggtcacaatagacaggggag cctcatgctt 1140 ataattctgc ttgccaagtg gaaatgctaa cctgctctttcccaggattt tagaaaaaca 1200 tttgaaatat ttcacaagat aaatgaaaat gggtaaacacaaggccttaa ggaaaatata 1260 cttctggtgc atttattaag cacttggatc tgatgcatgacacctgctta aaactcaaag 1320 aattatcaca ggctttgctg aggaagaaag aacaagaattatgactgcac acaggctcat 1380 acttcagcaa gtcaatgttt ttaaatcaat gaaaaacaatccagtgattg tgtttgctac 1440 aaagggaaag caatgtaaga tgacatacag tatttcaagctgttcaaatt actgatggca 1500 cagtggcaag atatatatcc attaaaacca aatagttgtttcaaagtatg ttggccaaca 1560 gagataaata atgtttctct gaagtgaata taaaataatgtctctcttca gaatgtagga 1620 ccctataatg gctttagcca caaatacctc ttgcttatttggggaatacg cattttccac 1680 cttcattcag aaattcttta gttagctcaa caaaattcatctttcacctc aacaaaaaag 1740 catgcgcatt aagtatcaaa gtatgtatat ttctacagtaggctcactga tggtctcatt 1800 cctattggta aatttgtcat agtgaatgag taaagtatgaataaatacaa tgtgttgaat 1860 gtagaggcta ctttcacaag aatattgcat catcagtgggccaatgttaa gagcatgttt 1920 gtgaggaaga ggaacataca atttctcttt ttttcttaattcttctctta tcagcatttt 1980 aattaataaa cataaaaagg taacactgat acacaaagtattaaatcacc atccattcgt 2040 ctttaaagat ttgcattctc ctaatcagtg aaaacaattctttctt 2086 51 941 DNA Homo sapiens 51 aaataaacat gctagctttt attccagttctaaccaaaaa aataaaccct cgttccacag 60 aagctgccat caagtatttc ctcacgcaagcaaccgcatc cataatcctt ctaatagcta 120 tcctcttcaa caatatactc tccggacaatgaaccataac caatactacc aatcaatact 180 catcattaat aatcataatg gctatagcaataaaactagg aatagccccc tttcacttct 240 gagtcccaga ggttacccaa ggcacccctctgacatccgg cctgcttctt ctcacatgac 300 aaaaactagc ccccatctca atcatataccaaatctctcc ctcactaaac gtaagccttc 360 tcctcactct ctcaatctta tccatcatagcaggcagttg aggtggatta aaccaaaccc 420 agctacgcaa aatcttagca tactcctcaattacccatat aggatgaata atagcagttc 480 taccgtacaa ccctaacata accattcttaatttaactat ttatattatc ctaactacta 540 ccgcattcct actactcaac ttaaactccagcaccacgac cctactacta tctcgcacct 600 gaaacaagct aacatgacta acacccttaattccatccac cctcctctcc ctaggaggcc 660 tgcccccgct aaccggcttt ttgcccaaatgggccattat cgaagaattc acaaaaaaca 720 atagcctcat catccccacc atcatagccaccatcaccct ccttaacctc tacttctacc 780 tacgcctaat ctactccacc tcaatcacactactccccat atctaacaac gtaaaaataa 840 aatgacagtt tgaacataca aaacccaccccattcctccc cacactcatc gcccttacca 900 cgctactcct acctatctcc ccttttatactaataatctt a 941 52 517 DNA Homo sapiens 52 ttaaaaatgc cctagcccacttcttaccac aaggcacacc tacacccctt atccccatac 60 tagttattat cgaaaccatcagcctactca ttcaaccaat agccctggcc gtacgcctaa 120 ccgctaacat tactgcaggccacctactca tgcacctaat tggaagcgcc accctagcaa 180 tatcaaccat taaccttccctctacactta tcatcttcac aattctaatt ctactgacta 240 tcctagaaat cgctgtcgccttaatccaag cctacgtttt cacacttcta gtaagcctct 300 acctgcacga caacacataatgacccacca atcacatgcc tatcatatag taaaacccag 360 cccatgaccc ctaacaggggccctctcagc cctcctaatg acctccggcc tagccatgtg 420 atttcacttc cactccataacgctcctcat actaggccta ctaaccaaca cactaaccat 480 ataccaatga tggcgcgatgtaacacgaga aagcaca 517 53 793 DNA Homo sapiens 53 ggcgaggcct atattacggatcatttctct actcagaaac ctgaaacatc ggcattatcc 60 tcctgcttgc aactatagcaacagccttca taggctatgt cctcccgtga ggccaaatat 120 cattctgagg ggccacagtaattacaaact tactatccgc catcccatac attgggacag 180 acctagttca atgaatctgaggaggctact cagtagacag tcccaccctc acacgattct 240 ttacctttca cttcatcttgcccttcatta ttgcagccct agcagcactc cacctcctat 300 tcttgcacga aacgggatcaaacaaccccc taggaatcac ctcccattcc gataaaatca 360 ccttccaccc ttactacacaatcaaagacg ccctcggctt acttctcttc cttctctcct 420 taatgacatt aacactattctcaccagacc tcctaggcga cccagacaat tataccctag 480 ccaacccctt aaacacccctccccacatca agcccgaatg atatttccta ttcgcctaca 540 caattctccg atccgtccctaacaaactag gaggcgtcct tgccctatta ctatccatcc 600 tcatcctagc aataatccccatcctccata tatccaaaca acaaagcata atatttcgcc 660 cactaagcca atcactttattgactcctag ccgcagacct cctcattcta acctgaatcg 720 gaggacaacc agtaagctacccttttacca tcattggaca agtagcatcc gtactatact 780 tcacaacaat cct 793 54827 DNA Homo sapiens 54 ggaaccttct ggttcctgcg ccaatcgcca ccagtatcaattgtgtgagc ttgggtgcga 60 gtgcacgcgt gcgtgagtac ggagagtata tatagatctctatctcttag caaaggtgaa 120 tgccagatgt aaatggcgcc tctgggcaaa ggaggcttgtattttgcaca ttttataaaa 180 acttgagaga atgagatttc tgcttgtata tttctaaaaagaggaaggag cccaaaccat 240 cctctcctta ccactcccat ccctgtgagc cctaccttacccctctgccc ctagccaagg 300 agtgtgaatt tatagatcta actttcatag gcaaaacaaaagcttcgagc tgttgcgtgt 360 gtgagtctgt tgtgtggatg tgcgtgtgtg gtccccagccccagactgga ttggaaaagt 420 gcatggtggg ggcctcgggg ctgtccccac gctgtccctttgccacaagt ctgtggggca 480 agaggctgca atattccgtc ctgggtgtct gggctgctaacctggcctgc tcaggcttcc 540 caccctgtgc ggggcacacc cccaggaagg gaccctggacacggctccca cgtccaggct 600 taaggtggat gcacttcccg cacctccagt cttctgtgtagcagctttaa cccacgtttg 660 tctgtcacgt ccagtcccga gacggctgag tgaccccaagaaaggcttcc ccgacaccca 720 gacagaggct gcagggctgg ggctgggtga gggtggcgggcctgcgggga cattctactg 780 tgctaaaaag ccactgcaga catagcaata aaaacatgtcattttcc 827 55 1562 DNA Homo sapiens 55 cccaccgact ccactatgtt gaagaaattcgacaagaagg atgaggagtc aggtggaggc 60 tccaacccat tccagcacct tgagaagagtgcggtactcc aggaggcccg tgtatttaat 120 gaaactccca tcaaccctcg gaaatgtgcccacatcctca ccaagattct ttatctcata 180 aaccaggggg agcacctggg gaccacggaagcgaccgagg ccttctttgc catgaccaag 240 ctctttcagt ccaatgatcc cacactccgtcggatgtgct acttgaccat caaggagatg 300 tcttgcattg cagaggatgt catcattgtcaccagcagcc taacaaaaga catgactggg 360 aaagaagaca actaccgggg cccggccgtgcgagccctct gccagatcac tgatagcacc 420 atgctgcagg ctattgagcg ctacatgaaacaagccattg tggacaaggt gcccagtgtc 480 tccagctctg ccctcgtgtc ttccttgcacctgctgaagt gcagctttga cgtggtcaag 540 cgctgggtga atgaggctca ggaggcagcatccagtgata acatcatggt ccagtaccac 600 gcactagggc tcctgtacca tgtgcgtaagaatgaccgcc tagccgtcaa taagatgatc 660 agcaaggtca cacggcatgg ccttaagtctccctttgcct actgcatgat gatccgggtg 720 gccagcaagc agctggaaga ggaggatggcagccgtgaca gcccactgtt tgacttcatc 780 gagagctgct tgcgcaacaa gcacgagatggtggtgtatg aagccgcctc ggccatcgtc 840 aatctgccag gctgcagtgc caaagagctggccccggctg tgtcagtgct ccagcttttc 900 tgcagctcac ccaaggctgc tctccgctatgctgctgttc gtaccctcaa taaggttgcc 960 atgaagcatc cgtcagctgt gacagcttgtaatctggatc tggagaacct ggtcacagat 1020 tcaaaccgca gcattgccac gctggccatcaccaccctcc ttaagacggg cagcgagagc 1080 agcatcgacc gcctcatgaa gcagatctcctccttcatgt cagaaatctc ggatgaattc 1140 aaggtggtgg ttgtccaggc catcagtgccctgtgtcaga aatatcctcg caaacacgcc 1200 gtccttatga acttcctgtt caccatgctgcgggaagagg gtggctttga gtataagcgc 1260 gctatcgtgg actgcatcat cagcatcattgaagagaact cagagagcaa ggagacaggg 1320 ctgtcacatc tgtgcgagtt catcgaggactgcgagttca cagtgctggc cacccgtatt 1380 ctacatctcc tgggccagga ggggcccaagaccaccaatc cctcaaagta catccgcttc 1440 atctataacc gagtggtctt ggagcatgaggaggtccggg caggtgctgt gagtgctctg 1500 gcgaagtttg gagcccagaa tgaagagatgttacccagta tcttggtgtt gctgaagagg 1560 tg 1562 56 678 DNA Homo sapiens 56ccccaaaccc actccacctt actaccagac aaccttagcc aaaccattta cccaaataaa 60gtataggcga tagaaattga aacctggcgc aatagatata gtaccgcaag ggaaagatga 120aaaattataa ccaagcataa tatagcaagg actaacccct ataccttctg cataatgaat 180taactagaaa taactttgca aggagagcca aagctaagac ccccgaaacc agacgagcta 240cctaagaaca gctaaaagag cacacccgtc tatgtagcaa aatagtggga agatttatag 300gtagaggcga caaacctacc gagcctggtg atagctggtt gtccaagata gaatcttagt 360tcaactttaa atttgcccac agaaccctct aaatcccctt gtaaatttaa ctgttagtcc 420aaagaggaac agctctttgg acactaggaa aaaaccttgt agagagagta aaaaatttaa 480cacccatagt aggcctaaaa gcagccacca attaagaaag cgttcaagct caacacccac 540tacctaaaaa atcccaaaca tataactgaa ctcctcacac ccaattggac caatctatca 600ccctatagaa gaactaatgt tagtataagt aacatgaaaa cattctcctc cgcataagcc 660tgcgtcagat taaaacac 678 57 1338 DNA Homo sapiens 57 tcacaacaagcaaacctgga gtatccttgg tctactccat gccctcccgg aacctgtccc 60 tgcggctggagggtctccag gagaaagact ctggccccta cagctgctcc gtgaatgtgc 120 aagacaaacaaggcaaatct aggggccaca gcatcaaaac cttagaactc aatgtactgg 180 ttcctccagctcctccatcc tgccgtctcc agggtgtgcc ccatgtgggg gcaaacgtga 240 ccctgagctgccagtctcca aggagtaagc ccgctgtcca ataccagtgg gatcggcagc 300 ttccatccttccagactttc tttgcaccag cattagatgt catccgtggg tctttaagcc 360 tcaccaacctttcgtcttcc atggctggag tctatgtctg caaggcccac aatgaggtgg 420 gcactgcccaatgtaatgtg acgctggaag tgagcacagg gcctggagct gcagtggttg 480 ctggagctgttgtgggtacc ctggttggac tggggttgct ggctgggctg gtcctcttgt 540 accaccgccggggcaaggcc ctggaggagc cagccaatga tatcaaggag gatgccattg 600 ctccccgaaccctgccctgg cccaagagct cagacacaat ctccaagaat gggacccttt 660 cctctgtcacctccgcacga gccctccggc caccccatgg ccctcccagg cctggtgcat 720 tgacccccacgcccagtctc tccagccagg ccctgccctc accaagactg cccacgacag 780 atggggcccaccctcaacca atatccccca tccctggtgg ggtttcttcc tctggcttga 840 gccgcatgggtgctgtgcct gtgatgatgc ctgcccagag tcaagctggc tctctggtat 900 gatgaccccaccactcattg gctaaaggat ttggggtctc tccttcctat aagggtcacc 960 tctagcacagaggcctgagt catgggaaag agtcacactc ctgaccctta gtactctgcc 1020 cccacctctctttactgtgg gaaaaccatc tcagtaagac ctaagtgtcc aggagacaga 1080 aggagaagaggaagtggatc tggaattggg aggagcctcc acccacccct gactcctcct 1140 tatgaagccagctgctgaaa ttagctactc accaagagtg aggggcagag acttccagtc 1200 actgagtctcccaggccccc ttgatctgta ccccacccct atctaacacc acccttggct 1260 cccactccagctccctgtat tgatataacc tgtcaggctg gcttggttag gttttactgg 1320 ggcagaggatagggaatc 1338 58 1999 DNA Homo sapiens 58 cccatttttg tatacctatcatccttccaa taaatcactc tttgcttagt ttaaccagac 60 tcagtttctg ttgctctcaactaaaacagt taacagataa tgattctata ttaaaatctc 120 ttagaatttc ttttctatatttttattggg tatttatcat gttagcagag cctgtaaact 180 ttaagggctt caagaaatttttaaaaaata tttcccggcc agatgcggtg gctcacacct 240 gtaatactag cactttgggaggccaaggcg ggcggatcac ctgaggtcag gagtttgaga 300 ccagcctgac caacatggagaaaccccatt tctactaaaa atacaaaatt agccgggcgt 360 ggtggtgcat gcctgtaatcccagctactc gggaggctga ggcaggagaa ttgcttgaac 420 ctgggaggcg gaggttgcggtgagtcaaga tcctgcaacc acattccggc ctgggcaaaa 480 aagcaagact ctttctcaaaaaaaaaaaaa aaaaaaaggt aaaatgacaa agtcatgcac 540 acacatgcta atctggaaagctataaatca aactgaaaac actggttatc ttgtagtgga 600 atggctggcc actttgactttctactttaa acatttggcc tatttattta ttttaaagag 660 acagggtctc actctgttgctgcccaagct ggagtagagt ggcgtaacca tagctcactg 720 taacctcaaa ctcctgggctcatgtaatcc tctcacttca gcctccagag tagctgagac 780 tatacaagtg cccgccaccatacctggtta attttttttt ttttttagat gaggtttccc 840 tatgttgccc aggctggtcttgaactcctg gcctcaagga atcctcccag tttggccttc 900 caaagtgctg gaattgcaggcataagccac catgcctggc ccagaacaaa cttattctta 960 tgtcatgttt gtggcaacaaaaattcaaaa aataatgaga cttccaggtt agctttaata 1020 gagagaggca agctagtgctttcacagtaa caactagata aaactaaata gattattcaa 1080 ataaataaat ggctttaaagattgaagagc tgtcttctat gcatagcaaa gaggactagc 1140 agaattaaaa ttccagagggagaagaggcc tctctaggtg agctgaaagt gccagccatt 1200 tcctctctct acttgtggtacttgtagagc tttagcgtga gtgatgataa ctggggcttg 1260 ccataggcag aactccatcaagagaaagaa aaaccagcag aacttatgac agctatatgg 1320 gttggcatga cagactaaatttcataacag ccctaaattt atggccagtt ttctacacag 1380 gacattcacc tccaggctggaggcctgaac tagaaaacca aagtgaaatt aagcacaatc 1440 tcacagtgct tagcagataaagctagaggg agtggcctgt aacaaacata caacggatcc 1500 cactcttggg atactacaccgaactgaagg acttaaatct tcagcccagt ccaactcagt 1560 tgtttggata gaggtgttttatacctcatt gtatctgcct aagactgcaa aatgcaaaca 1620 ttaactttag actctaaaggtctttacaca cattgtctag aaatcgtcaa aaacttctgc 1680 tgcctggcgc agtggctcacgtctgtagtt ccagcatttt gggaggccga ggcaggctga 1740 tcatgaggtc aggacttcgagaccagcctg accaacatgg tgaaaccctc tctctactga 1800 aaatacaaaa atttgccaggcatggtggcg cacgcccata atcccagcta ctggggaggc 1860 tgaggcagga gagccgcttgaacccaggag gcaaaggttg cagtgagcca aaaccgcgtg 1920 ccactgcact ccagcctgggcaacagagcg agactctgtc tcaaaaaaca acaacaactt 1980 ttgctttccc aggagggag1999 59 957 DNA Homo sapiens 59 ggcctgctca aggtggtgtt cgtggtcttcgcctccttgt gtgcctggta ttcggggtac 60 ctgctcgcag agctcattcc agatgcacccctgtccagtg ctgcctatag catccgcagc 120 atcggggaga ggcctgtcct caaaggtgagtgccgtgctt ggggcagacg gctgccttca 180 tggcttgtgt gcagagggag gagtggtggtttctgtccca gctggaggct ggggggccct 240 gacggcttta tatcaggaag gaggaggagagaagcatttt gcagttactc atttaggtcc 300 cacgccattc caaggcacgt gccgctgctgaccccatgtg agactagaac agtgtggctt 360 ctggttacca gcacaaacct gcagccagaccttgaactgt gctctttccg cccagatctt 420 ggcttcctcc cctgggaagc acaggagggtccaggttctg agatcccgaa tcattttaag 480 gtttctgtcg gactgaagag ctgttggaaaatcttctgta aagtcttggg aagcaaaagc 540 tagtaattga gtagatgtga aagaaagggcagacattgta tgtcggtgaa cacacggcct 600 caaccatcag cttaggaaag gcccagctctctgtagggcg gctgctttct gtccttcccc 660 acaatcaagg tgggttttct gagagcctgctgtgtgctgg gcatgtccag gagctaagga 720 gagacggagg acaagcccac gaggtccctgcccctttggc actgataccc tagtgagggg 780 aggtgacagc ttaacgcatg aacaaatgagattcgagaga ggatgggtgg ggaaagcctc 840 tgagaaggag gtgacaattg agaagcatctcagtgaagca agggagtggg ccatgctggg 900 ttctgagggc tgtgtgccag tttgggagcatcatgactta gtctacccag attactg 957 60 941 DNA Homo sapiens 60 cttaggaaactggagaggat attcatggtc ttaagcacta gattatagag tttaggttgc 60 cagagtaacagtttcaaaga tcctggcaca cacagttgct ctttagtaaa tagttcttaa 120 ataaattgagacatgtacat ttattctcaa tttcctcttt cttatttgtt ttccagagtg 180 tttccttactctcagaagtt taaaaaatta agtacttggg gctgggcgca gtgactcacg 240 cctgtaatcccagcactttg ggaggctgag gtggccagat cataaggtca agagatcgag 300 atcatcctggccaacatggt gaaaccccgt ctctactaaa aatacaaaaa ctagctgagc 360 gtggtggcaggcacctgtag tcccagctat ttgggaggct gaggcagcag aatcacttga 420 acccaggaggggaggttgca gtgagccgag atcgcgccat gcacaccagc ctgggtgata 480 gagcgagactccatctccaa aaaaaaaaaa agtacttggg tacagtggct cacacctgta 540 atcctagcatttggggagac caaggtggga ggatcacttg agcctgggag gtcaaggctg 600 cagtgagacatgtttgagca ctgcattcca acctgggtga cagagagcct atcttattgt 660 tgtataaaggtttgaaatag gtacaaggtt tggtcattca gtttcaaatt catttctcaa 720 atatccttgtatctcactaa catcatctgt ctaccagtaa catttccaaa tctaatagta 780 cttggcaagagatggggagg gggttaaatt gcacaattta aaattttcat aattgttatg 840 catcacagttattgtgatca cgtgataatg ttatgcatgt cttttgttgt gtgtcagccc 900 caacttttgtctcagcattt gaaccatgca tacatcttat c 941 61 2131 DNA Homo sapiens 61agaagctgcc atgacctgaa ttaggtaatg ggtagtggac agaagggcac aactgcccat 60gcctgggagg gaacaaaggg gtgttgactc ctgtgaaccc acttcccaag ctctatatca 120gagctcctgg gctgcatagg agtggccagg ctaacaacct gaccccttcc cttcctccct 180gggcagctcc ctgcggtgtg gccccccaag cacgcatcac aggtggcagc agtgcagtcg 240ccggtcagtg gccctggcag gtcagcatca cctatgaagg cgtccatgtg tgtggtggct 300ctctcgtgtc tgagcagtgg gtgctgtcag ctgctcactg cttccccagg taccaaatgg 360tgaaagtcaa aagtgggctg gaggtcaaag ttaatgggtc aatccaggtc agaggttggg 420gttcttgggt tgggctgaga gtgtcagttc taagtctgga aggaccagtt aggagtgaag 480gtgaggggtc agaggttaca tgataaaact aaggtgcaat ttgagaccta aaaggggctg 540atcggaatca ggggttggca gtgggggttg gggaacaagg agaggtctcc tgggtctcag 600cctttgcccc ctgcctgcag cgagcaccac aaggaagcct atgaggtcaa gctgggggcc 660caccagctag actcctactc cgaggacgcc aaggtcagca ccctgaagga catcatcccc 720caccccagct acctccagga gggctcccag ggcgacattg cactcctcca actcagcaga 780cccatcacct tctcccgcta catccggccc atctgcctcc ctgcagccaa cgcctccttc 840cccaacggcc tccactgcac tgtcactggc tggggtcatg tggccccctc aggtgaggtg 900ggacgtgggt gcctagaggt gtggaggggc acctgacttg gggaggggcc cagggtaagc 960ctcttttacc cccacagtga gcctcctgac gcccaagcca ctgcagcaac tcgaggtgcc 1020tctgatcagt cgtgagacgt gtaactgcct gtacaacatc gacgccaagc ctgaggagcc 1080gcactttgtc caagaggaca tggtgtgtgc tggctatgtg gaggggggca aggacgcctg 1140ccaggtaagc acaggcccgg gggcagatga ccagtgcaac ttcggaaagg aggcctggcc 1200cggtcctgat ggctgctgtg tggcttctct cctcagggtg actctggggg cccactctcc 1260tgccctgtgg agggtctctg gtacctgacg ggcattgtga gctggggaga tgcctgtggg 1320gcccgcaaca ggcctggtgt gtacactctg gcctccagct atgcctcctg gatccaaagc 1380aaggtgacag aactccagcc tcgtgtggtg ccccaaaccc aggagtccca gcccgacagc 1440aacctctgtg gcagccacct ggccttcagc tctgccccag cccagggctt gctgaggccc 1500atccttttcc tgcctctggg cctggctctg ggcctcctct ccccatggct cagcgagcac 1560tgagctggcc ctacttccag gatggatgca tcacactcaa ggacaggagc ctggtccttc 1620cctgatggcc tttggaccca gggcctgact tgagccactc cttccttcag gactctgcgg 1680gaggctgggg ccccatcttg atctttgagc ccattcttct gggtgtgctt tttgggacca 1740tcactgagag tcaggagttt tactgcctgt agcaatggcc agagcctctg gcccctcacc 1800caccatggac cagcccattg gccgagctcc tggggagctc ctgggaccct tggctatgaa 1860aatgagccct ggctcccacc tgtttctgga agactgctcc cggcccgctg cccagactga 1920tgagcacatc tctctgccct ctccctgtgt tctgggctgg ggcacctttg tgcagcttcg 1980aggacaggaa aggccccaat cttgcccact ggccgctgag cgcccccgag ccctgactcc 2040tggactccgg aggactgagc ccccaccgga actgggctgg cgcttggatc tggggtggga 2100gtaacagggc agaaatgatt aaaatgtttg a 2131 62 1032 DNA Homo sapiens 62ctacgcggcc gagatctgca tcgccctcaa cttcctgcac gagaggggga tcatctacag 60ggacctgaag ctggacaacg tcctcctgga tgcggacggg cacatcaagc tcacagacta 120cggcatgtgc aaggaaggcc tgggccctgg tgacacaacg agcactttct gcggaacccc 180gaattacatc gcccccgaaa tcctgcgggg agaggagtac gggttcagcg tggactggtg 240ggcgctggga gtcctcatgt ttgagatgat ggccgggcgc tccccgttcg acatcatcac 300cgacaacccg gacatgaaca cagaggacta ccttttccaa gtgatcctgg agaagcccat 360ccggatcccc cggttcctgt ccgtcaaagc ctcccatgtt ttaaaaggat ttttaaataa 420ggaccccaaa gagaggctcg gctgccggcc acagactgga ttttctgaca tcaagtccca 480cgcgttcttc cgcagcatag actgggactt gctggagaag aagcaggcgc tccctccatt 540ccagccacag atcacagacg actacggtct ggacaacttt gacacacagt tcaccagcga 600gcccgtgcag ctgaccccag acgatgagga tgccataaag aggatcgacc agtcagagtt 660cgaaggcttt gagtatatca acccattatt gctgtccacc gaggagtcgg tgtgaggccg 720cgtgcgtctc tgtcgtggac acgcgtgatt gaccctttaa ctgtatcctt aaccaccgca 780tatgcatgcc aggctgggca cggctccgag ggcggccagg gacagacgct tgcgccgaga 840ccgcagaggg aagcgtcagc gggcgctgct gggagcagaa cagtccctca cacctgggcc 900cgggcaggcc agcttcgtgc tggaggaact tgctgctgtg cctgcgtcgc ggcggatccg 960cggggaccct gccgaggggg ctgtcatgcg gtttccaagg tgcacatttt ccacggaaac 1020agaactcgat gc 1032 63 615 DNA Homo sapiens 63 gtttctggat actgccttcaatctccaggc ctttgagggc aagacatttt agagctgggc 60 aaaacctgtg taggtctcgctgtgggtttg ctggggacca agggggtgat gaaaagggga 120 ggggcggagc tcctgcccaagagaggggct gtggggcccc aggataaaac agacacagtg 180 acagggccaa gagccagcactgctggcctt ggtgtcatgc cagaatctac caggactgag 240 ggagccagag gagtcctgtaggcaggctac tgtgctggag catcccccag ctgctcccat 300 cttgctggaa tttcttgggcggcttctcca cctgtatctc aagacagaca cccgggggcc 360 tgtgtctgtg gccgctcccatcccggcagc cctggctgct gctcgcccca ccctcgctta 420 tctgtagatt caaagcgatgttctcttctg tgctcttaga agtagggagt tcagcagtaa 480 cagccaggtg aagcgaacctgctgggtgat ttgtttgcgc tctgttttat ggggcattcc 540 tgcgagatgt gtcagcttctgtatgagatg cagccacagc tcatgtgtac caaagtagaa 600 aaccaaatca cagag 615 64123 DNA Artificial Sequence Description of Artificial Sequenceprimer 64aggcggcaac gccgaggaga ggagaggaag gccccagaaa accaggagga agaggaggag 60cgtgcagaac tgaatcagtc ggaggaacct gaggcaggcg agagtagtac tggagggcct 120tga 123 65 29 DNA Artificial Sequence Description of ArtificialSequenceprimer 65 acgtgaattc aggcggcaac gccgaggag 29 66 32 DNAArtificial Sequence Description of Artificial Sequenceprimer 66cgatctcgag tcaaggccct ccagtactac tc 32 67 72 PRT Homo sapiens 67 Lys CysGlu Arg Val Arg Ser Trp Val Asn Phe Asp Ser Ser Arg Leu 1 5 10 15 ProGly Ser Leu Arg Ala Ser Ser Ser Ser Arg Ser Pro Val Ala Ala 20 25 30 GlySer Val Arg Arg Ser Ala Pro Ala Leu Ala Leu Pro Trp Ala Leu 35 40 45 ProAla Pro Pro Arg Trp Pro Ala Ser Ala Ser Gln Pro Gly Pro Gly 50 55 60 ProGly Gly Ser Gly Ser Pro Arg 65 70 68 13 PRT Homo sapiens 68 Ser Thr MetLeu Ala Gly Leu Ala Ala Leu Phe Leu Ala 1 5 10 69 25 PRT Homo sapiens 69Val Asn Tyr Ala Gly Trp Thr Gly Cys Leu Val Pro Gly Leu Gly Leu 1 5 1015 Ser Phe Ile Thr Ile Thr Cys Thr Asp 20 25 70 35 PRT Homo sapiens 70Lys Asp Ser Leu Phe Asn Lys Trp Cys Trp Glu Asn Trp Ile Ser Ile 1 5 1015 Cys Lys Arg Met Lys Leu Asp Leu Tyr Leu Thr Pro Leu Ala Lys Ile 20 2530 Ser Ser Lys 35 71 11 PRT Homo sapiens 71 Gln Pro Arg Pro Pro Ile TyrSer Ser His Leu 1 5 10 72 49 PRT Homo sapiens 72 Ile Asn Met Leu Ala PheIle Pro Val Leu Thr Lys Lys Ile Asn Pro 1 5 10 15 Arg Ser Thr Glu AlaAla Ile Lys Tyr Phe Leu Thr Gln Ala Thr Ala 20 25 30 Ser Ile Ile Leu LeuIle Ala Ile Leu Phe Asn Asn Ile Leu Ser Gly 35 40 45 Gln 73 25 PRT Homosapiens 73 Gly Glu Ala Tyr Ile Thr Asp His Phe Ser Thr Gln Lys Pro GluThr 1 5 10 15 Ser Ala Leu Ser Ser Cys Leu Gln Leu 20 25 74 13 PRT Homosapiens 74 Arg Gly Leu Tyr Tyr Gly Ser Phe Leu Tyr Ser Glu Thr 1 5 10 7537 PRT Homo sapiens 75 Ser Gln Gln Ala Asn Leu Glu Tyr Pro Trp Ser ThrPro Cys Pro Pro 1 5 10 15 Gly Thr Cys Pro Cys Gly Trp Arg Val Ser ArgArg Lys Thr Leu Ala 20 25 30 Pro Thr Ala Ala Pro 35 76 59 PRT Homosapiens 76 Thr Pro Thr Pro Ser Leu Ser Ser Gln Ala Leu Pro Ser Pro ArgLeu 1 5 10 15 Pro Thr Thr Asp Gly Ala His Pro Gln Pro Ile Ser Pro IlePro Gly 20 25 30 Gly Val Ser Ser Ser Gly Leu Ser Arg Met Gly Ala Val ProVal Met 35 40 45 Met Pro Ala Gln Ser Gln Ala Gly Ser Leu Val 50 55 77 19PRT Homo sapiens 77 Lys Tyr Lys Asn Leu Pro Gly Met Val Ala His Ala HisAsn Pro Ser 1 5 10 15 Tyr Trp Gly 78 109 PRT Homo sapiens 78 Ala Cys SerArg Trp Cys Ser Trp Ser Ser Pro Pro Cys Val Pro Gly 1 5 10 15 Ile ArgGly Thr Cys Ser Gln Ser Ser Phe Gln Met His Pro Cys Pro 20 25 30 Val LeuPro Ile Ala Ser Ala Ala Ser Gly Arg Gly Leu Ser Ser Lys 35 40 45 Val SerAla Val Leu Gly Ala Asp Gly Cys Leu His Gly Leu Cys Ala 50 55 60 Glu GlyGly Val Val Val Ser Val Pro Ala Gly Gly Trp Gly Ala Leu 65 70 75 80 ThrAla Leu Tyr Gln Glu Gly Gly Gly Glu Lys His Phe Ala Val Thr 85 90 95 HisLeu Gly Pro Thr Pro Phe Gln Gly Thr Cys Arg Cys 100 105 79 35 PRT Homosapiens 79 Pro Ala Gln Gly Gly Val Arg Gly Leu Arg Leu Leu Val Cys LeuVal 1 5 10 15 Phe Gly Val Pro Ala Arg Arg Ala His Ser Arg Cys Thr ProVal Gln 20 25 30 Cys Cys Leu 35 80 60 PRT Homo sapiens 80 Lys Val LysLeu Leu Arg Ser Leu Pro Gln Arg Phe Lys Met Asp Val 1 5 10 15 His IleThr Pro Gly Thr His Ala Ser Glu His Ala Val Asn Lys Gln 20 25 30 Leu AlaAsp Lys Glu Arg Val Ala Ala Ala Leu Glu Asn Thr His Leu 35 40 45 Leu GluVal Val Asn Gln Cys Leu Ser Ala Arg Ser 50 55 60 81 35 PRT Homo sapiens81 Gln Asn Ile His Leu Ile Arg Ala Pro Leu Ala Gly Lys Gly Lys Gln 1 510 15 Leu Glu Glu Lys Met Val Gln Gln Leu Gln Glu Asp Val Asp Met Glu 2025 30 Asp Ala Pro 35 82 118 PRT Homo sapiens 82 Phe Lys Pro Lys Pro ProLys Asn Glu Ser Leu Glu Thr Tyr Pro Val 1 5 10 15 Met Lys Tyr Asn ProAsn Val Leu Pro Val Gln Cys Thr Gly Lys Arg 20 25 30 Asp Glu Asp Lys AspLys Val Gly Asn Val Glu Tyr Phe Gly Leu Gly 35 40 45 Asn Ser Pro Gly PhePro Leu Gln Tyr Tyr Pro Tyr Tyr Gly Lys Leu 50 55 60 Leu Gln Pro Lys TyrLeu Gln Pro Leu Leu Ala Val Gln Phe Thr Asn 65 70 75 80 Leu Thr Met AspThr Glu Ile Arg Ile Glu Cys Lys Ala Tyr Gly Glu 85 90 95 Asn Ile Gly TyrSer Glu Lys Asp Arg Phe Gln Gly Arg Phe Asp Val 100 105 110 Lys Ile GluVal Lys Ser 115 83 98 PRT Homo sapiens 83 Asn Glu Ser Glu Val Lys GlyTyr Lys Val Leu Tyr Arg Trp Asn Arg 1 5 10 15 Gln Ser Ser Thr Ser ValIle Glu Thr Asn Lys Thr Ser Val Glu Leu 20 25 30 Ser Leu Pro Phe Asp GluAsp Tyr Ile Ile Glu Ile Lys Pro Phe Ser 35 40 45 Asp Gly Gly Asp Gly SerSer Ser Glu Gln Ile Arg Ile Pro Lys Ile 50 55 60 Ser Asn Ala Tyr Ala ArgGly Ser Gly Ala Ser Thr Ser Asn Ala Cys 65 70 75 80 Thr Leu Ser Ala IleSer Thr Ile Met Ile Ser Leu Thr Ala Arg Ser 85 90 95 Ser Leu 84 147 PRTHomo sapiens 84 Gly Ile Gln Ser Thr Pro Leu Asn Leu Ala Val Asn Trp ArgCys Glu 1 5 10 15 Pro Ser Ser Thr Asp Leu Arg Ile Asp Tyr Lys Tyr AsnThr Asp Ala 20 25 30 Met Thr Thr Ala Val Ala Leu Asn Asn Val Gln Phe LeuVal Pro Ile 35 40 45 Asp Gly Gly Val Thr Lys Leu Gln Ala Val Leu Pro ProAla Val Trp 50 55 60 Asn Ala Glu Gln Gln Arg Ile Leu Trp Lys Ile Pro AspIle Ser Gln 65 70 75 80 Lys Ser Glu Asn Gly Gly Val Gly Ser Leu Leu AlaArg Phe Gln Leu 85 90 95 Ser Glu Gly Pro Ser Lys Pro Ser Pro Leu Val ValGln Phe Thr Ser 100 105 110 Glu Gly Ser Thr Leu Ser Gly Cys Asp Ile GluLeu Val Gly Ala Gly 115 120 125 Tyr Arg Phe Ser Leu Ile Lys Lys Arg PheAla Ala Gly Lys Tyr Leu 130 135 140 Ala Asp Asn 145 85 76 PRT Homosapiens 85 Met Ser Lys Ala His Pro Pro Glu Leu Lys Lys Phe Met Asp LysLys 1 5 10 15 Leu Ser Leu Lys Leu Asn Gly Gly Arg His Val Gln Gly IleLeu Arg 20 25 30 Gly Phe Asp Pro Phe Met Asn Leu Val Ile Asp Glu Cys ValGlu Met 35 40 45 Ala Thr Ser Gly Gln Gln Asn Asn Ile Gly Met Val Val IleArg Gly 50 55 60 Asn Ser Ile Ile Met Leu Glu Ala Leu Glu Arg Val 65 7075 86 240 PRT Homo sapiens 86 Pro Asp Pro Arg Pro Ser Pro Pro Arg ProAsp Val Cys Met Ala Asp 1 5 10 15 Pro Glu Gly Leu Ser Ser Glu Ser GlyArg Val Glu Arg Leu Arg Glu 20 25 30 Lys Glu Lys Val Gln Gly Arg Val GlyArg Arg Ala Pro Gly Lys Ala 35 40 45 Lys Pro Ala Ser Pro Ala Arg Arg LeuAsp Leu Arg Gly Lys Arg Ser 50 55 60 Pro Thr Pro Gly Lys Gly Pro Ala AspArg Ala Ser Arg Ala Pro Pro 65 70 75 80 Arg Pro Arg Ser Thr Thr Ser GlnVal Thr Pro Ala Glu Glu Lys Asp 85 90 95 Gly His Ser Pro Met Ser Lys GlyLeu Val Asn Gly Leu Lys Ala Gly 100 105 110 Pro Met Ala Leu Ser Ser LysGly Ser Ser Gly Ala Pro Val Tyr Val 115 120 125 Asp Leu Ala Tyr Ile ProAsn His Cys Ser Gly Lys Thr Ala Asp Leu 130 135 140 Asp Phe Phe Arg ArgVal Arg Ala Ser Tyr Tyr Val Val Ser Gly Asn 145 150 155 160 Asp Pro AlaAsn Gly Glu Pro Ser Arg Ala Val Leu Asp Ala Leu Leu 165 170 175 Glu GlyLys Ala Gln Trp Gly Glu Asn Leu Gln Val Lys Val Thr Leu 180 185 190 IlePro Thr His Asp Thr Glu Val Thr Arg Glu Trp Tyr Gln Gln Thr 195 200 205His Glu Gln Gln Gln Gln Leu Asn Val Leu Val Leu Ala Ser Ser Ser 210 215220 Thr Val Val Met Gln Asp Glu Ser Phe Pro Ala Cys Lys Ile Glu Phe 225230 235 240 87 183 DNA Homo sapiens 87 aaggtcaagc ttctgcgctc ccttcctcagcgtttcaaga tggacgtgca cattactccg 60 gggacccatg cctcagagca tgcagtgaacaagcaacttg cagataagga gcgggtggca 120 gctgccctgg agaacaccca cctcttggaggttgtgaatc agtgcctgtc agcccgctcc 180 tga 183 88 108 DNA Homo sapiens 88caaaacatcc atcttatccg agcccctctt gcaggcaaag ggaaacagtt ggaagagaaa 60atggtacagc agttacaaga ggatgtggac atggaagatg ctccttaa 108 89 357 DNA Homosapiens 89 ttcaaaccta agcctcccaa gaatgagtcc ttggagactt acccagtgatgaagtataac 60 ccaaatgtcc ttcccgttca gtgcactggc aagcgagatg aagataaggataaagttgga 120 aatgtggagt attttggact gggcaactcc cctggttttc ctctgcagtattatccgtac 180 tatggcaaac tcctgcagcc caaatacctg cagcccctgc tggccgtacagttcaccaat 240 cttaccatgg acactgaaat tcgcatagag tgtaaggcgt acggtgagaacattgggtac 300 agtgagaaag accgttttca gggacgtttt gatgtaaaaa ttgaagttaagagctga 357 90 297 DNA Homo sapiens 90 aatgagtcgg aagtaaaagg atacaaagtcttgtacagat ggaacagaca aagcagcaca 60 tctgtcattg aaacaaataa aacatcggtggagctttctt tgcctttcga tgaagattat 120 ataatagaaa ttaagccatt cagcgacggaggagatggca gcagcagtga acaaattcga 180 attccaaaga tatcaaatgc ctacgcgagaggatctgggg cttccacttc gaatgcatgt 240 acgctgtcag ccatcagtac aataatgatttccctcacag ctaggtccag tttatga 297 91 444 DNA Homo sapiens 91 ggcattcagtccacacctct gaacctggca gtgaattggc gatgtgagcc ttcaagcact 60 gacctgcgcatagattacaa atataataca gatgcaatga cgactgctgt ggccctcaac 120 aatgtgcagttcctggtccc catcgacgga ggagtcacca agctccaggc agtgctccca 180 ccagcagtctggaatgctga acaacagaga atattgtgga agattcctga tatctctcag 240 aagtcagaaaatggaggggt gggttctttg ttggcaagat ttcagttatc tgaaggccca 300 agcaaaccttctccattggt tgtgcagttc acaagtgaag gaagcaccct ttctggctgt 360 gacattgaacttgttggagc agggtatcga ttttcactca tcaagaaaag gtttgctgca 420 ggaaaatacttggcagataa ctaa 444 92 231 DNA Homo sapiens 92 atgagcaaag ctcaccctcccgagttgaaa aaatttatgg acaagaagtt atcattgaaa 60 ttaaatggtg gcagacatgtccaaggaata ttgcggggat ttgatccctt tatgaacctt 120 gtgatagatg aatgtgtggagatggcgact agtggacaac agaacaatat tggaatggtg 180 gtaatacgag gaaatagtatcatcatgtta gaagccttgg aacgagtata a 231 93 723 DNA Homo sapiens 93ccagaccccc gcccatcccc tccccgccct gatgtgtgca tggctgaccc cgaggggctc 60agctcagagt ctgggagagt agagaggcta cgggagaagg aaaaggttca ggggcgagta 120gggcgcaggg ccccaggcaa ggccaagcca gcgtcccctg cacggcgtct ggatcttcgg 180ggaaaacgct cacccacccc tggtaaaggg cctgcagatc gagcatcccg ggccccacct 240cgaccacgca gcaccacaag ccaggtcacc ccagcagagg aaaaggatgg acacagcccc 300atgtccaaag gcctagtcaa tggactcaag gcaggaccaa tggccttgag ttccaagggc 360agctctggtg cccctgtata tgtggatctc gcctacatcc cgaatcattg cagtggcaag 420actgctgacc ttgacttctt ccgtcgagtg cgtgcatcct actatgtggt cagtgggaat 480gaccctgcca atggcgagcc aagccgggct gtgctggatg ccctgctgga gggcaaggcc 540cagtgggggg agaatcttca ggtgaaggtg actctgatcc ctactcatga cacggaggtg 600actcgtgagt ggtaccaaca aactcatgag cagcagcaac aactgaatgt cctggtcctg 660gctagcagca gcaccgtggt gatgcaggat gagtccttcc ctgcctgcaa gattgagttc 720tga 723

1. A polypeptide, which is a substantially purified polypeptide, whereinthe polypeptide has following characteristics: (a) the polypeptidedirectly or indirectly binds to a cytoplasmic domain of the receptor foran advanced glycation endproduct (AGE); and (b) the polypeptide inhibitsa signal transduction from binding of a ligand to the receptor foradvanced glycation endproduct (AGE) through activation of NFκB.
 2. Thepolypeptide according to claim 1, which comprises the amino acidsequence selected from SEQ ID NOS: 1 to 32 and SEQ ID NOS: 67 to 79 inthe Sequence Listing or said amino acid sequence including deletion,substitution, or addition of one or several amino acids.
 3. Thepolypeptide according to claim 1, which comprises the amino acidsequence selected from SEQ ID NOS: 11, 12, 29, and 30 in the SequenceListing or said amino acid sequence including deletion, substitution, oraddition of one or several amino acids.
 4. The polypeptide according toclaim 1, which comprises the amino acid sequence selected from SEQ IDNOS: 80 to 86 in the Sequence Listing or said amino acid sequenceincluding deletion, substitution, or addition of one or several aminoacids.
 5. A polynucleotide, which encodes the polypeptide according toclaim
 1. 6. The polynucleotide according to claim 5, comprising thenucleotide sequence represented by one selected from SEQ ID NOS: 33 to63 or a nucleotide sequence which is hybridizable with thepolynucleotide represented by one selected from SEQ ID NOS: 33 to 63under a stringent condition.
 7. The polynucleotide according to claim 5,comprising the nucleotide sequence represented by one selected from SEQID NOS: 43, 44, 61, and 62 or a nucleotide sequence which ishybridizable with the polynucleotide represented by one selected fromSEQ ID NOS: 43, 44, 61, and 62 under a stringent condition.
 8. Thepolynucleotide according to claim 5, comprising the nucleotide sequencerepresented by one selected from SEQ ID NOS: 87 to 93 or a nucleotidesequence which is hybridizable with the polynucleotide represented byone selected from SEQ ID NOS: 87 to 93 under a stringent condition.
 9. Apharmaceutical for a gene therapy, comprising the polynucleotideaccording to claim 5 and a vector which is expressible in an animal. 10.The pharmaceutical for a gene therapy according to claim 9, which isused for treating a disease selected from diabetes, a diabeticcomplication, Alzheimer's Disease, dialysis amyloidosis, a cancer, aperiodontal disease, and an aging-related disease.
 11. A vector,comprising the polynucleotide according to claim
 5. 12. A microorganismor cell, which is transformed with the vector according to claim
 11. 13.A method for producing a polypeptide comprising: culturing amicroorganism or cell which is transformed with a vector comprising thepolynucleotide according to claim 5; and isolating the polypeptideencoded by the polynucleotide from the culture.
 14. A method forscreening a substance that inhibits or accelerates the biding of thepolypeptide according to claim 1 to a cytoplasmic domain of the receptorfor advanced glycation endproduct (AGE), comprising: placing a target ofscreening in a screening system that contains the cytoplasmic domain ofthe receptor for advanced glycation endproduct (AGE) and the polypeptideand measuring a degree of inhibition or acceleration of the binding ofthe polypeptide to a cytoplasmic domain of the receptor for advancedglycation end product (AGE).
 15. A method for screening a substance thatenhances or inhibits a function of the polypeptide according to claim 1or a substance that increases or decreases an amount of the polypeptidecomprising: placing a target of screening in a screening system thatcontains the polypeptide and measuring a degree of enhancement orinhibition of the function of the polypeptide or a degree of increase ordecrease of the amount of the polypeptide.
 16. A compound, which isobtainable by the screening method according to claim
 14. 17. Thecompound according to claim 16, which has following characteristics: (a)the compound directly or indirectly binds to a cytoplasmic domain of thereceptor for advanced glycation endproduct (AGE); and (b) the compoundinhibits a signal transduction from binding of a ligand to the receptorfor advanced glycation endproduct (AGE) through activation of NFκB. 18.A pharmaceutical composition, comprising the substance selected from thegroup consisting of the compound according to claim 16, a salt thereof,a hydrate thereof, and a solvate thereof.
 19. The pharmaceuticalcomposition according to claim 18, which is used for treating a diseaseselected from diabetes, a diabetic complication, Alzheimer's Disease,dialysis amyloidosis, a cancer, a periodontal disease, and anaging-related disease.
 20. An antibody, which can specifically bind tothe polypeptide according to claim
 1. 21. A polynucleotide, which isrepresented by a nucleotide sequence having at least 20 sequentialnucleotides in any of the sequences of SEQ ID NOS: 33 to 63 in theSequence Listing, a sequence complementary to the nucleotide sequence,or a nucleotide sequence which is hybridizable with the nucleotidesequence under a stringent condition.
 22. A polynucleotide, which isrepresented by a nucleotide sequence having at least 20 sequentialnucleotides in any of the sequences of SEQ ID NOS: 43, 44, 61, and 62 inthe Sequence Listing, a sequence complimentary to the nucleotidesequence, or a nucleotide sequence which is hybridizable with thenucleotide sequence under a stringent condition.
 23. A polynucleotide,which is represented by a nucleotide sequence having at least 20sequential nucleotides in any of the sequences of SEQ ID NOS: 87 to 93in the Sequence Listing, a sequence complimentary to the nucleotidesequence, and a nucleotide sequence which is hybridizable with thenucleotide sequence under a stringent condition.
 24. A probe, comprisinga label and the polynucleotide according to claim
 21. 25. A reagent fordiagnosis, comprising the probe according to claim
 24. 26. The reagentfor diagnosis according to claim 25, which is used for a diagnosis ofdiabetes, a diabetic complication, Alzheimer's Disease, dialysisamyloidosis, a cancer, a periodontal disease, and an aging-relateddisease.